Forecasting carbon futures price: a hybrid method incorporating fuzzy entropy and extreme learning machine

In this paper, we propose a novel hybrid model that extends prior work involving ensemble empirical mode decomposition (EEMD) by using fuzzy entropy and extreme learning machine (ELM) methods. We demonstrate this 3-stage model by applying it to forecast carbon futures prices which are characterized by chaos and complexity. First, we employ the EEMD method to decompose carbon futures prices into a couple of intrinsic mode functions (IMFs) and one residue. Second, the fuzzy entropy and K-means clustering methods are used to reconstruct the IMFs and the residue to obtain three reconstructed components, specifically a high frequency series, a low frequency series, and a trend series. Third, the ARMA model is implemented for the stationary high and low frequency series, while the extreme learning machine (ELM) model is utilized for the non-stationary trend series. Finally, all the component forecasts are aggregated to form final forecasts of the carbon price for each model. The empirical results show that the proposed reconstruction algorithm can bring more than 40% improvement in prediction accuracy compared to the traditional fine-to-coarse reconstruction algorithm under the same forecasting framework. The hybrid forecasting model proposed in this paper also well captures the direction of the price changes, with strong and robust forecasting ability, which is significantly better than the single forecasting models and the other hybrid forecasting models.</p

Supplement Number 1

The supplementary material has three sections, which are coupling coefficient between two neighboring cavities, measured experimental data, and stimulated results for higher frequency

table_2_Pituitary Action of E2 in Prepubertal Grass Carp: Receptor Specificity and Signal Transduction for Luteinizing Hormone and Follicle-Stimulating Hormone Regulation.docx

<p>17β-estradiol (E2) is an important sex steroid produced by ovary and brain. In mammals, E2 plays an important role in hypothalamus–pituitary–gonad axis to regulate puberty onset, however, little is known about the functional role of E2 in teleost pituitary. Using prepubertal grass carp as model, three nuclear estrogen receptors (nERs: estrogen receptor alpha, estrogen receptor beta 1, and estrogen receptor beta 2) and two G protein-coupled estrogen receptors (GPER1: GPER1a and GPER1b) were isolated from grass carp pituitary. Tissue distribution analysis indicated that both nERs and GPERs were highly detected in grass carp pituitary, which suggested that E2 should play an important role in grass carp pituitary. Using primary cultured grass carp pituitary cells as model, high-throughput RNA-seq was used to examine the E2-induced differentially expressed genes (DEGs). Transcriptomic analysis showed that E2 could significantly upregulate the expression of 28 genes in grass carp pituitary cells, which were characterized into different functions including reproduction, gonad development, and central nervous system development. Further studies confirmed that E2 could induce luteinizing hormone (LH) and follicle-stimulating hormone (FSH) secretion and mRNA expression in prepubertal grass carp pituitary in vivo and in vitro. In the pituitary, LH and FSH regulation by E2 were mediated by both ERβ and GPER1. Apparently, E2-induced LHβ and FSHβ mRNA expression were mediated by adenylyl cyclase/cAMP/protein kinase A, phospholipase C/inositol 1,4,5-triphosphate/protein kinase C, and Ca²⁺/calmodulin/CaM-dependent protein kinase II pathways. In addition to LH and FSH, E2 could also induce growth regulation by estrogen in breast cancer 1 (a novel regulator for pituitary development) mRNA expression in grass carp pituitary cells. These results, as a whole, suggested that E2 could play an important role in gonadotropin hormone release and pituitary development in prepubertal grass carp.</p

table_1_Pituitary Action of E2 in Prepubertal Grass Carp: Receptor Specificity and Signal Transduction for Luteinizing Hormone and Follicle-Stimulating Hormone Regulation.docx

<p>17β-estradiol (E2) is an important sex steroid produced by ovary and brain. In mammals, E2 plays an important role in hypothalamus–pituitary–gonad axis to regulate puberty onset, however, little is known about the functional role of E2 in teleost pituitary. Using prepubertal grass carp as model, three nuclear estrogen receptors (nERs: estrogen receptor alpha, estrogen receptor beta 1, and estrogen receptor beta 2) and two G protein-coupled estrogen receptors (GPER1: GPER1a and GPER1b) were isolated from grass carp pituitary. Tissue distribution analysis indicated that both nERs and GPERs were highly detected in grass carp pituitary, which suggested that E2 should play an important role in grass carp pituitary. Using primary cultured grass carp pituitary cells as model, high-throughput RNA-seq was used to examine the E2-induced differentially expressed genes (DEGs). Transcriptomic analysis showed that E2 could significantly upregulate the expression of 28 genes in grass carp pituitary cells, which were characterized into different functions including reproduction, gonad development, and central nervous system development. Further studies confirmed that E2 could induce luteinizing hormone (LH) and follicle-stimulating hormone (FSH) secretion and mRNA expression in prepubertal grass carp pituitary in vivo and in vitro. In the pituitary, LH and FSH regulation by E2 were mediated by both ERβ and GPER1. Apparently, E2-induced LHβ and FSHβ mRNA expression were mediated by adenylyl cyclase/cAMP/protein kinase A, phospholipase C/inositol 1,4,5-triphosphate/protein kinase C, and Ca²⁺/calmodulin/CaM-dependent protein kinase II pathways. In addition to LH and FSH, E2 could also induce growth regulation by estrogen in breast cancer 1 (a novel regulator for pituitary development) mRNA expression in grass carp pituitary cells. These results, as a whole, suggested that E2 could play an important role in gonadotropin hormone release and pituitary development in prepubertal grass carp.</p

Synthesis, Biological Evaluation, and Autophagy Mechanism of 12<i>N</i>‑Substituted Sophoridinamines as Novel Anticancer Agents

A series of 12<i>N</i>-substituted sophoridinamine derivatives were synthesized and evaluated for their cytotoxic activities in human HepG2 hepatoma cells. Structure–activity relationship revealed that introduction of a suitable arylidene or arylethyl at the <i>N</i>′-end could greatly enhance antiproliferation potency. Among them, compound <b>6b</b> possessing a <i>N</i>′-trimethoxyphenyl methylene exhibited potent antiproliferation effect against three human tumor cell lines including HepG2, leukemia (K562), and breast cancer (HMLE), with IC₅₀ between 0.55 and 1.7 μM. The underlying mechanism of <b>6b</b> against tumor cells is to block autophagic flux, mainly through neutralizing lysosomal acidity. Our results indicated that compound <b>6b</b> is a potent lysosomal deacidification agent and is accordingly able to block autophagic flux and inhibit tumor cell growth

Pharmaceutical inhibition of p38α reduces adiposity and enhances the browning of WAT.

<p>(A and B) Representative HE staining of iBAT, iWAT, and eWAT (A), diameter and cross-sectional area (B) of adipocytes in these adipose tissues from SB203580-treated C57BL/6J mice at 2 d postinjection. Bars: 100 μm. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (C) BW of C57BL/6J mice before and after 4 wk of SB203580 treatment (control: <i>n</i> = 4, SB203580: <i>n</i> = 5). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (D and E) Relative weight of iWAT and eWAT from C57BL/6J mice after 4 wk of SB203580 treatment. These mice were maintained at RT (D, <i>n</i> = 5 per group) or exposed to cold for 2 d before analysis (E, <i>n</i> = 5 per group) as indicated. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (F) Relative mRNA levels of UCP-1, PGC1α, ELOVL3, and DIO2 in iWAT from C57BL/6J mice after 4 wk of SB203580 treatment (<i>n</i> = 10 per group). These mice were exposed to cold for 2 d before analysis. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (G) Representative western blots of p-CREB (Ser133) and UCP-1 in iWAT from C57BL/6J mice after 4 wk of SB203580 treatment. These mice were exposed to cold for 2 d before analysis. (H and I) Representative PET/CT images (H) and PET images (I) of C57BL/6J mice after 4 wk of SB203580 treatment. These mice received a daily CL316,243 injection for 8 d before ¹⁸F-FDG injection. White dashed triangles represent the anatomical sites of iWAT. (J) Ex vivo measured ¹⁸F-FDG uptake in iBAT, iWAT, and eWAT to tissue weight ratio by γ counter (<i>n</i> = 3 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (K) Relative weight of iWAT from <i>db/db</i> mice after 3 wk of SB203580 treatment (<i>n</i> = 5 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (L and M) Representative HE staining of iWAT (L), diameter and cross-sectional area of adipocytes in iWAT (M) from <i>db/db</i> mice after 3 wk of SB203580 treatment. Bars: 100 μm. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. Means ± SEM are shown. *<i>p</i> < 0.05; **<i>p</i> < 0.01; ***<i>p</i> < 0.001. BW, body weight; CREB, cAMP-response element binding protein; CT, computed tomography; DIO2, deiodinase 2; ELVOL3, elongation of very long chain fatty acids (FEN1/Elo2, SUR4/Elo3, yeast)-like 3; eWAT, epididymal white adipose tissue; HE staining, hematoxylin-eosin staining; iBAT, interscapular brown adipose tissue; iWAT, inguinal white adipose tissue; NS, not significant; PET, positron emission tomography; PGC1α, peroxisome proliferative activated receptor gamma coactivator 1α; RT, room temperature; UCP-1, uncoupling protein 1; WAT, white adipose tissue.</p

Adipocyte-specific deletion of p38α leads to a lean phenotype and increased glucose tolerance and insulin sensitivity.

<p>(A-C) Representative western blots of p38α and p-p38 in iBAT (A), iWAT (B), and eWAT (C) from Floxed and Fp38αKO mice as indicated. (D-F) Representative western blots of p38α in other tissues, including liver (D), skeletal muscle (E), and macrophages (F) from Floxed and Fp38αKO mice as indicated. (G) Cumulative gross energy intake and feces of Floxed and Fp38αKO mice for 24 h (<i>n</i> = 4 per group). Mice were maintained at RT. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (H) Growth curve of Floxed (<i>n</i> = 15) and Fp38αKO (<i>n</i> = 8–13) mice maintained at RT. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (I and J) BW, FM, and FM to BW ratio (FM/BW) of Floxed and Fp38αKO mice maintained at RT (<i>n</i> = 11 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (K and L) GTT (K, <i>n</i> = 8 per group) and ITT (L, <i>n</i> = 8 per group) in Floxed and Fp38αKO mice. AUCs were calculated. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. Means ± SEM are shown. *<i>p</i> < 0.05; **<i>p</i> < 0.01. AUC, area under curve; BW, body weight; eWAT, epididymal white adipose tissue; FM, fat mass; GTT, glucose tolerance test; iBAT, interscapular brown adipose tissue; ITT, insulin tolearance test; iWAT, inguinal white adipose tissue; NS, not significant; RT, room temperature.</p

Ablation of p38α in adipose tissues facilitates the browning of WAT.

<p>(A and B) Representative HE staining of iWAT (A), diameter and cross-sectional area of adipocytes in iWAT (B) from Floxed and Fp38αKO mice exposed to cold for 2 d. Bars: 100 μm. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (C) Relative mRNA levels of UCP-1, PGC1α, ELOVL3, DIO2, COX8B, and CIDEAin iWAT from Floxed and Fp38αKO mice maintained at RT or exposed to cold for 2 d (RT, <i>n</i> = 4 per group; Cold, <i>n</i> = 8–16 per group). Fold of change was indicated (Floxed versus Fp38αKO). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (D) Representative western blots of UCP-1 and TH in iWAT from Floxed and Fp38αKO mice maintained at RT or exposed to cold for 2 d. (E) Representative UCP-1 staining of iWAT from Floxed and Fp38αKO mice exposed to cold for 2 d. Bars: 100 μm. (F) Relative mRNA levels of CD137, TBX1, and TMEM26 in iWAT from Floxed (<i>n</i> = 3) and Fp38αKO (<i>n</i> = 6) mice exposed to cold for 2 d. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (G) Relative mitDNA to nuDNA ratio in unilateral iWAT of Floxed and Fp38αKO mice exposed to cold for 2 d (<i>n</i> = 6 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (H) Representative western blots of p38α, p-p38, p-ATF2, and UCP-1 in iWAT from Floxed and Fp38αKO mice exposed to cold for 2 d. (I) Relative p-ATF2 protein levels in iWAT of Floxed and Fp38αKO mice exposed to cold for 2 d. The densities of p-ATF2 bands were quantitated and normalized to Hsp90 (<i>n</i> = 3 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (J and K) VO₂ (J) and VCO₂ (K) within 48 h after CL316,243 injection in Floxed (<i>n</i> = 5) and Fp38αKO (<i>n</i> = 4) mice maintained at RT. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (L and M) Representative HE staining of iWAT (L), diameter and cross-sectional area of adipocytes in iWAT (M) from Floxed and Fp38αKO mice exposed to cold for 7 d. Bars: 100 μm. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (N) Relative mRNA levels of UCP-1, PGC1α, PRDM16, ELOVL3, DIO2, COX8B, and CIDEA in iWAT from Floxed and Fp38αKO mice exposed to cold for 7 d (<i>n</i> = 4 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (O) Representative UCP-1 staining of iWAT from Floxed and Fp38αKO mice exposed to cold for 7 d. Bars: 100 μm. (P) Relative mRNA levels of UCP-1, COX8B, DIO2, ELOVL3, PGC1α, and PRDM16 in iWAT from 5-wk-old Floxed (<i>n</i> = 6) and Fp38αKO (<i>n</i> = 6) mice maintained at RT. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (Q–S) Total amounts of protein in iBAT (<i>n</i> = 6–9) and iWAT (<i>n</i> = 5–11), and total UCP-1 protein per depot (<i>n</i> = 3) in both iBAT and iWAT of Floxed and Fp38αKO mice maintained at RT or exposed to cold for 2 d. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. Means ± SEM are shown. *<i>p</i> < 0.05; **<i>p</i> < 0.01; ***<i>p</i> < 0.001. ATF2, activating transcription factor 2; ATGL, adipose triglyceride lipase; BAT, brown adipose tissue; BT, body temperature; BW, body weight; CIDEA, cell death-inducing DNA fragmentation factor, alpha subunit-like effector A; COX8B, cytochrome c oxidase subunit 8B; DIO2, deiodinase 2; ELVOL3, elongation of very long chain fatty acids (FEN1/Elo2, SUR4/Elo3, yeast)-like 3; HE staining, hematoxylin-eosin staining; HSL, hormone-sensitive lipase; iBAT, interscapular brown adipose tissue; IHC, immunohistochemistry; iWAT, inguinal white adipose tissue; MGL, monoglyceride lipase; mitDNA, mitochondrial DNA; ND, not detectable; NS, not significant; nuDNA, nuclear DNA; OCR, oxygen consumption rate; PGC1α, peroxisome proliferative activated receptor gamma coactivator 1α; PRDM16, positive regulatory domain containing 16; RT, room temperature; TBX1, T-box 1; TH, tyrosine hydroxylase; TMEM26, transmembrane protein 26; UCP-1, uncoupling protein 1; VCO₂, carbon dioxide production; VO₂, oxygen consumption; WAT, white adipose tissue.</p

The effect of p38α inhibition or deficiency on WAT browning is cell autonomous.

<p>(A-C) Representative HE staining of iWAT (A), diameter and cross-sectional area of adipocytes in iWAT (B), and representative UCP-1 staining of iWAT (C) from C57BL/6J mice after injection of Ad-p38αAF into the fat pad of iWAT. These mice were exposed to cold for 2 d before analysis. Bars: 100 μm. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (D-F) Relative mRNA levels of UCP-1, PGC1α, DIO2, COX8B, and/or ELVOL3 in iWAT-SVF-derived matured adipocytes treated with SB203580 for 4 or 8 h (D, <i>n</i> = 4 per group), or p38α-specific inhibitor (p38αMAPK-IN-1) for 4 h (E, <i>n</i> = 4 per group), or infected with Lenti-p38αAF (F, <i>n</i> = 3 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (G) Relative mRNA levels of UCP-1, PGC1α, PRDM16, DIO2, ELVOL3, COX8B, and CIDEA in matured adipocytes derived from iWAT-SVF of Floxed and Fp38αKO mice (<i>n</i> = 4 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (H) Representative fluorescence staining of Mito in matured iWAT-SVF-derived adipocytes infected with Lenti-p38αAF. Bars: 25 μm. (I and J) OCR of Oligomycin, FCCP, and Antimycin A/Rotenone-treated matured adipocytes derived from iWAT-SVF of Floxed and Fp38αKO mice (I) and the AUC of OCR (J) as indicated (<i>n</i> = 5 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (K and L) Representative UCP-1 staining of iWAT-SVF-derived matured adipocytes after infection with Lenti-p38αAF (K) or matured adipocytes derived from iWAT-SVF of Floxed and Fp38αKO mice (L). Bars: 10 μm. Means ± SEM are shown. *<i>p</i> < 0.05; **<i>p</i> < 0.01; ***<i>p</i> < 0.001. Ad-p38αAF, adenovirus expressing p38αAF; AUC, area under the curve; CIDEA, cell death-inducing DNA fragmentation factor, alpha subunit-like effector A; COX8B, cytochrome c oxidase subunit 8B; DIO2, deiodinase 2; ELVOL3, elongation of very long chain fatty acids (FEN1/Elo2, SUR4/Elo3, yeast)-like 3; FCCP, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; HE staining, hematoxylin-eosin staining; IHC, immunohistochemistry; iWAT, inguinal white adipose tissue; Lenti-p38αAF, lentivirus expressing p38αAF; MAPK, mitogen-activated protein kinase; Mito, mitochondria; NS, not significant; OCR, oxygen consumption rate; PGC1α, peroxisome proliferative activated receptor gamma coactivator 1α; PRDM16, positive regulatory domain containing 16; SVF, stromal vascular fraction; UCP-1, uncoupling protein 1; WAT, white adipose tissue.</p

Loss of p38α in adipose tissues causes minimal effects on BAT.

<p>(A) BT of Floxed and Fp38αKO mice maintained at RT (<i>n</i> = 6 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (B and C) VO₂ (B) and VCO₂ (C) in Floxed and Fp38αKO mice maintained at RT (<i>n</i> = 4 per group). The values were normalized by LM. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (D) iBAT weight and relative iBAT weight to BW ratio (iBAT/BW) of Floxed (<i>n</i> = 10) and Fp38αKO (<i>n</i> = 8). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (E-G) Representative HE staining of iBAT (E), diameter and cross-sectional area of adipocytes in iBAT (F), and representative UCP-1 staining of iBAT (G) from Floxed and Fp38αKO mice maintained at RT. Bars: 100 μm. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (H) Representative EM images of iBAT from Floxed and Fp38αKO mice maintained at RT at low (top), medium (middle), and high (bottom) magnification, as indicated. (I) Relative mitDNA to nuDNA ratio in unilateral iBAT of Floxed (<i>n</i> = 5) and Fp38αKO (<i>n</i> = 9) mice maintained at RT. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (J) OCR of ADP, Oligomycin, FCCP, and Antimycin A/Rotenone-treated mitochondria derived from iBAT of Floxed and Fp38αKO mice exposed to cold for 2 d (<i>n</i> = 4 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (K) Relative mRNA levels of UCP-1, PGC1α, DIO2, COX8B, and PRDM16 in iBAT from Floxed and Fp38αKO mice maintained at RT (<i>n</i> = 6 per group) or exposed to cold for 2 d (<i>n</i> = 8 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (L and M) Representative western blots of UCP-1 in iBAT from Floxed and Fp38αKO mice maintained at RT (L) or exposed to cold for 2 d (M). (N) BT of Fp38αKO and Floxed mice exposed to cold for 4 h (<i>n</i> = 4 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (O) Relative mRNA levels of UCP-1 and PGC1α in iBAT from Floxed (<i>n</i> = 7–8) and Fp38αKO (<i>n</i> = 8) mice exposed to cold for 4 h. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (P and Q) Representative western blots (P) and densitometry analysis (Q) of UCP-1 in iBAT from Floxed and Fp38αKO mice exposed to cold for 4 h. The densities of UCP-1 bands were quantitated and normalized to Hsp90 (<i>n</i> = 4 per group). See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. (R and S) Relative mRNA levels of PRDM16, DIO2, ELVOL3, COX8B, and CIDEA(R), ATGL, MGL, and HSL (S) in iBAT from Floxed (<i>n</i> = 7–8) and Fp38αKO (<i>n</i> = 6–8) mice exposed to cold for 4 h. See also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004225#pbio.2004225.s010" target="_blank">S1 Data</a>. Means ± SEM are shown. *<i>p</i> < 0.05; **<i>p</i> < 0.01; ***<i>p</i> < 0.001. ADP, adenosine diphosphate; ATGL, adipose triglyceride lipase; BAT, brown adipose tissue; BT, body temperature; BW, body weight; CIDEA, cell death-inducing DNA fragmentation factor, alpha subunit-like effector A; COX8B, cytochrome c oxidase subunit 8B; DIO2, deiodinase 2; ELVOL3, elongation of very long chain fatty acids (FEN1/Elo2, SUR4/Elo3, yeast)-like 3; EM, electron microscopy; FCCP, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; HE staining, hematoxylin-eosin staining; HSL, hormone-sensitive lipase; iBAT, interscapular brown adipose tissue; IHC, immunohistochemistry; LM, lean mass; MGL, monoglyceride lipase; mitDNA, mitochondrial DNA; NS, not significant; nuDNA, nuclear DNA; OCR, oxygen consumption rate; PGC1α, peroxisome proliferative activated receptor gamma coactivator 1α; PRDM16, positive regulatory domain containing 16; RT, room temperature; UCP-1, uncoupling protein 1; VCO₂, carbon dioxide production; VO₂, oxygen consumption.</p