Figure 7.

Prmt5 regulates fatty acid metabolic pathways through interacting with SREBP1a. a) Heatmap of top 25 significant changed TAGs from eWAT of WT (Prmt5^flox/flox) and Prmt5^AKO mice (upper) and list of TAGs that were decrease by >2‐fold in Prmt5^AKO compared to WT (Prmt5^flox/flox) eWAT (lower), n = 5 pairs male mice of 6‐month‐old. b) Concentration of FFA, cholesterol, HDL, LDL, and TAG from the serum of WT (Prmt5^flox/flox) and Prmt5^AKO mice, n = 4 pairs male mice of 6‐month‐old. c) Heatmap of top 25 significant changed TAGs from WT (Prmt5^flox/flox) and Prmt5^AKO mice (upper) and list of TAGs that were decreased by >1.6‐fold in Prmt5^AKO compared to WT (Prmt5^flox/flox) eWAT (lower), after 12‐week of HFD, n = 4 pairs of male mice. d) Concentration of FFA, cholesterol, HDL, LDL, and TAG from the serum of WT (Prmt5^flox/flox) and Prmt5^AKO mice, n = 3 pairs male mice after 12‐week HFD feeding. e) Fold change of Free Fatty Acids (FFA) in eWAT of WT (Prmt5^flox/flox) and Prmt5^AKO mice. f) Ratio of 18:1/16:0 FFA indicating de novo lipogenesis. e,f) n = 5 pairs of 6‐month‐old male mice. g,h) Expression of g) adipogenic and h) lipogenic genes in eWAT from WT (Prmt5^flox/flox) and Prmt5^AKO mice, n = 3 pairs of male mice at 6‐month‐old. i) Representative Western blots showing relative protein levels of PPARγ, ATGL, CEBPα, FABP4, and β‐Tubulin from eWAT of male WT (Prmt5^flox/flox) and Prmt5^AKO mice after 12‐week of HFD. j) Heatmap from RNA‐sequencing data showing relative expression of genes related to Fatty acid transport, TAG synthesis and LD biogenesis in eWAT of WT (Prmt5^flox/flox) and Prmt5^AKO mice, n = 3 pairs male mice of 6‐month‐old. k) A diagramatic pathway showing enzymes involved in triacylglycerol (TAG) synthesis, red arrows represent genes down‐regulated in Prmt5^AKO eWAT. i) Lysates from 293T cells overexpressing Flag‐SRBEP1a or Flag‐SRBEP1a and Myc‐PRMT were immunoprecipitated (IP) with Flag or PRMT5 antibody and blotted with Flag, PRMT5, SYM10, and GAPDH antibodies. m) Representative Western blots showing relative protein levels of SREBP1, PRMT5, and β‐Tubulin from eWAT of male WT (Prmt5^flox/flox) and Prmt5^AKO mice after 12 week of HFD. n) Representative Western blots showing relative protein levels of SREBP1, PRMT5, GAPDH, and β‐Tubulin from control and PRMT5‐overexpressed 3T3‐L1 cell line. o) Expression of genes related to TAG synthesis after PRMT5 overexpression, samples were collected from three different wells with three technical repeats for each. p) Representative Western blots showing the overexpression of Flag‐SRBEP1a in SVF preadipocytes from iWAT of 8‐week‐old male WT (Prmt5^flox/flox) and Prmt5^AKO mice after 8 day of differentiation in vitro. q) Expression of Bscl2, Fasn, and Dgat1 in SVF preadipocytes from iWAT of 8‐week‐old male WT (Prmt5^flox/flox) and Prmt5^AKO mice with/without Flag‐SRBEP1a overexpression after 8 day of differentiation in vitro, n = 3 pairs of mice and cells from each mouse received three individual transfections. Data represent mean ± s.e.m. (t‐test: *p < 0.05, **p < 0.01).