The reliability of colorimetry is precise(ly) as expected.

Figure S6. Gene ontology (GO) analysis of the differentially expressed genes between C1 and C2. (JPG 403 kb

Dietary dibutyryl cAMP supplementation regulates the fat deposition in adipose tissues of finishing pigs via cAMP/PKA pathway

This study investigated potential mechanism of dibutyryl-cAMP (db-cAMP) on porcine fat deposition. (1) Exp.1, 72 finishing pigs were allotted to 3 treatments (0, 10 or 20 mg/kg dbcAMP) with 6 replicates. dbcAMP increased the hormone sensitive lipase (HSL) activity and expression of β-adrenergic receptor (β-AR) and growth hormone receptor (GHR), but decreased expression of peroxisome proliferator-activated receptor gamma 2 (PPAR-γ2) and adipocyte fatty acid binding protein (A-FABP) in back fat. dbcAMP upregulated expression of β-AR, GHR, PPAR-γ2 and A-FABP, but decreased insulin receptor (INSR) expression in abdominal fat. Dietary dbcAMP increased HSL activity and expression of G protein-coupled receptor (GPCR), cAMP-response element-binding protein (CREB) and insulin-like growth factor-1 (IGF-1), but decreased fatty acid synthase (FAS) and lipoprotein lipase (LPL) activities, and expression of INSR, cAMP-response element-binding protein (C/EBP-α) and A-FABP in perirenal fat. (2) Exp. 2, dbcAMP suppressed the proliferation and differentiation of porcine preadipocytes in a time- and dose-dependent manner, which might be associated with increased activities of cAMP and protein kinase A (PKA), and expression of GPCR, β-AR, GHR and CREB via inhibiting C/EBP-α and PPAR-γ2 expression. Collectively, dbcAMP treatment may reduce fat deposition by regulating gene expression related to adipocyte differentiation and fat metabolism partially via cAMP-PKA pathway.</p

Table_1_Low Protein Diet Improves Meat Quality and Modulates the Composition of Gut Microbiota in Finishing Pigs.DOCX

This study investigated the effect of a low protein (LP) diet on growth performance, nitrogen emission, carcass traits, meat quality, and gut microbiota in finishing pigs. Fifty-four barrows (Duroc × Landrace × Yorkshire) were randomly assigned to three treatments with six replicates (pens) of three pigs each. The pigs were fed with either high protein (HP, 16% CP), medium protein (MP, 12% CP), and LP diets (10% CP), respectively. The LP diets did not influence the growth performance, but significantly decreased the plasma urea nitrogen contents and fecal nitrogen emission (P * (lightness) value of meat color, and muscle fiber density in the longissimus dorsi (P * value at 24 and 48 h, and a significant negative correlation between unidentified_Ruminococcasceae in both ileum and colon with L* value at 24 h (P < 0.05). Collectively, the LP diet supplemented with lysine, methionine, threonine, and tryptophan could reduce the fecal nitrogen emission without affecting growth performance and improve meat quality by regulating the antioxidant capacity and gene expression involved in fat metabolism as well as modulating the gut microbiota composition in finishing pigs.</p

Additional file 1 of Genetic modifications of critical regulators provide new insights into regulation modes of raw-starch-digesting enzyme expression in Penicillium

Additional file 1: Fig. S1. Southern blot analysis of the genomic DNA of OamyR, OhepA, ΔCreA, and Mpga3 mutants. The location of the probes and restriction enzyme sites for Southern blot analysis are shown. The primers are listed in Table S2. (A) The overexpression strains OamyR and OhepA generated 5.6 and 4.7 kb DNA bands, respectively, while the parental strain 114–2 did not produce a detectable band, indicating that the overexpression cassettes were integrated into the genome. (B) Southern blot analysis of the genomic DNA of 114-2 and Mpga3. The 114-2 strain generated a 1.3 kb DNA band, while a 3.1 kb band was obtained in Mpga3 indicating that the targeted gene was correctly replaced. Sequencing of the mutated pga3 (Mpga3) gene in strain Mpga3. The mutated codon is indicated. (C) Southern blot analysis of the genomic DNA of 114-2 and ΔCreA. The 114-2 strain generated a 3.3 kb DNA band, while a 6.3 kb band was obtained in ΔCreA indicating that the targeted gene was correctly replaced. Fig. S2. RSDE activity assay of WT and various mutants. The strains were cultivated in liquid VMM supplemented with 1% starch and cultivated at 30 °C for 5 days. Soluble starch was used as a reaction substrate for RSDE activity assays. Fig. S3. RSDE activity assay of WT, ΔHepA and ΔPGA3 strain. The strains were cultivated in liquid VMM supplemented with 1% starch and cultivated at 30 °C for 5 days. Raw rice starch was used as a reaction substrate for RSDE activity assays. Fig. S4. Extracellular proteins of 114-2 on starch analysed by SDS-PAGE. 32 μL culture supernatant of 114-2 strain was loaded after cultivating in 1% starch medium for 72 h, respectively. Lane M was the molecular weight marker, lanes 1 and 2 represent two independent cultivations for 114-2 strain. Fig. S5. SDS-PAGE analysis of the purified raw starch digesting enzyme Amy15A. Lane M indicates protein molecular weight marker; lane 1 indicates the purified Amy15A. Fig. S6. Effects of pH and temperature on enzymatic activity and the stability of the RSDE Amy15A. (A) The effect of pH on enzyme activity. The enzyme activity was assayed in a citrate–phosphate buffer (pH 3.0–7.0) at 37 °C. (B) The pH stability of Amy15A was measured by pre-incubating the enzyme in various buffers for 24 h at 4 °C, and the residual enzyme activity was determined using the standard method. (C) The influence of temperature on enzyme activity. The enzyme activity was determined between 35 °C and 80 °C under optimum pH condition. (D) The influence of temperature on enzyme stability. Temperature stability was determined by the standard method after pre-incubating the enzyme at pH 4.5 between 30 °C and 75 °C for 1 h. Data given are mean ± standard deviation from three replicates. The results are from a representative experiment, and similar results were obtained in two other independent experiments. Table S1. Quality control statistics. Table S2. Primers used in this study

Additional file 11: of Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells

Figure S9. Expression of adult Leydig cell (ALC)-associated markers in induced differentiated C2 clusters. They did not express the PLC markers PDGFRA1 (a) and LIFR (b), but expressed testosterone synthesis enzymes CYP11A1 (c), CYP17A1 (d), and StAR (e), demonstrated at both the protein and mRNA levels (f). (JPG 301 kb

Additional file 6: of Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells

Figure S4. Expression of pluripotency-associated markers and germ cell-specific markers in the C1 and C2 clusters. Immunofluorescent staining showed that both the C1 and C2 clusters expressed the pluripotency-associated markers SSEA1, SSEA4, TRA-1-60, and TRA-1-81. However, the C1 clusters, but not the C2 clusters, expressed the germ cell-specific markers GFRA1 and PLZF. (JPG 249 kb

Additional file 9: of Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells

Figure S7. Cell cycle assay of putative porcine progenitor Leydig cells (PLCs). Cell cycle analysis of the PLCs showed that they had rapidly dividing capacities at days 3, 5, and 7 in culture. Analysis of apoptosis showed that PLCs had very low levels of cell death at day 7 in culture (99.81% propidium iodide, annexin V double negative). (JPG 299 kb

Additional file 7: of Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells

Figure S5. Comparison of pluripotency potential between the C1 clusters and porcine-induced pluripotent stem cells (piPSCs). (a) The C1 clusters and (b) piPSCs were cultured without feeder cells and serum for 7 days to induce embryoid body formation. Embryoid bodies were formed from the piPSCs, but not the C1 clusters, (c) with induction of lineage-specific genes. (JPG 310 kb

Additional file 3: of Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells

Figure S1. Enrichment of porcine male germline stem cell (mGSCs). (a) The testicular suspension contained several cell types. (b) Testicular fibroblast cells (TFCs) were removed by differential attachment technique. (c) The unattached cells underwent density gradient centrifuging to remove the leukomonocytes (LYMs) and cell debris (d) and red blood cells (RBCs) (e), and to retain mGSCs (f). (JPG 699 kb

Additional file 1: of Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells

Table S1. Primary and secondary antibodies used for immunofluorescence or FACS analysis. (DOCX 15 kb