Fig. 4: IGFBP1 mediates the induction of lipolysis and NASH in Wtap-HKO mice.
From: Deficiency of WTAP in hepatocytes induces lipoatrophy and non-alcoholic steatohepatitis (NASH)
a Heatmap of top differentiated genes encoding secreted proteins in the livers of Wtapflox/flox and Wtap-HKO mice (Wtap-HKO fpkm > 500 and FoldChange > 4.5). Data were presented as log 10 fpkm (n = 3 for each group). b Relative Igfbp1 mRNA levels in the liver and eWAT (n = 8 for each group; Liver, P = 0.0014; eWAT, P = 0.547). c IGFBP1 protein levels in the liver and eWAT were measured by immunoblotting (n = 4 for each group). The samples were derived from the same experiment and the blots were processed in parallel. d Serum IGFBP1 protein levels were measured by ELISA (n = 11 for each group; P = 0.0011). e–p Wtap-HKO mice at 7 weeks old were intravenously injected with a control antibody (IgG) and an anti-IGFBP1 neutralizing antibody per day, respectively, for 5 days. Serum FFA levels were measured (e) (IgG, n = 7; Anti-IGFBP1, n = 8; P = 0.0455). p-HSL, HSL, p-PKA substrate, ATGL and GAPDH protein levels in eWAT were measured by immunoblotting and quantified by Image J (f) (n = 3 for each group; p-HSL/HSL, P = 0.0445; ATGL/GAPDH, P = 0.0335; p-PKA substrate/GAPDH, P = 0.0499). cAMP levels were measured by ELISA (g) (IgG, n = 7; Anti-IGFBP1, n = 8; P = 0.00998). The ADCY3, ADCY4 and ADCY6 protein levels in the eWAT were measured by immunoblotting and quantified by Image J (h) (n = 3 for each group; ADCY3, P = 0.0009; ADCY4, P = 0.0318; ADCY6, P = 0.00918). serum ALT activity were measured (i) (n = 6 for each group; P = 0.0031). Cleaved caspase 3 levels (j) (n = 3 for each group; P = 0.0012), liver TAG levels (k) (n = 6 for each group; P < 0.0001), hepatic lipid droplets (l), TUNEL-positive cells (m, n) (n = 5 for each group; P = 0.0129), and cytochemokine mRNA levels (o) (n = 5 for each group; Tnfα, P = 0.0294; Il1β, P = 0.0133; Il6, P = 0.0267; iNos, P = 0.3452; Infg, P = 0.6684; Cd14, P = 0.0111; Csf1, P = 0.0292; Ccl2, P = 0.0172; Ccl3, P = 0.0824; Ccl5, P = 0.0025; Ccl22, P = 0.0722; Ccr2, P = 0.0969; Cxcl2, P = 0.0693; Cxcl5, P = 0.092; Cxcl10, P = 0.0747; Cx3cl1, P = 0.1067) were measured in the liver. Serum cytokine levels were measured by ELISA (p) (IgG, n = 7; Anti-IGFBP1, n = 8; TNFα, P = 0.000178; IL1β, P = 0.0323; CCL2, P = 0.0456). For immunoblotting, the samples were derived from the same experiment and the blots were processed in parallel. n was the number of biologically independent mice. Data represent the mean ± SEM. Significance was determined by unpaired two-tailed Student’s t test analysis. *p < 0.05. **p < 0.01. Source data are provided as a Source Data file.
