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

Abstract

<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