Figs2.

Supplementary Figure 2. The densitometry analysis of Western blots. Image Lab was used to analysis the densitometry of Western blotting bands of AKT1+2 + 3, p-AKT, GSK-3β, p-GSK-3β, FOXO1 and p-FOXO1 in MPH cells (A). Relative gray scale was calculated by dividing the relative gray values (i.e., AKT1+2 + 3/β-ACTIN) in “∗∗” p < 0.01, Ad-B4 vs. Ad-RFP, “^##” p < 0.01, Ad-siB4 vs. Ad-RFP. Image Lab was used to analysis the densitometry of Western blotting bands of AKT1+2 + 3, p-AKT, GSK-3β, p-GSK-3β, FOXO1 and p-FOXO1 in 4 week liver tissue (B) and 12 week liver tissue (C). Relative gray scale was calculated by dividing the relative gray values (i.e., AKT1+2 + 3/β-ACTIN) in “∗∗” p < 0.01, “∗” p < 0.05, Ad-B4 vs. Ad-GFP. The effect of BMP4 on hepatic glucose concentrations of HFD mice in vivo. Recombinant adenovirus Ad-B4 or Ad-GFP were injected in liver area of 4-week old C57BL/6 mice (male, n = 10 /time point/group), which were fed up with 45% high fat diet (HFD). The mice were sacrificed after 4 weeks and 12 weeks, respectively. The liver sections were subjected to H & E staining and PAS staining (D), which observed by light microscopic examination (x400) and the serum glucose levels were also measured (E), and total RNA was isolated from the liver of Ad-B4 HFD group and Ad-GFP HFD group mice, and TqPCR analysis was carried out to detect the expression of the genes that govern glycogen synthesis, gluconeogenesis and glucose metabolism. Relative expression was calculated by dividing the relative expression values (i.e., gene/Gapdh) in “∗∗” p < 0.01, Ad-B4 HFD group vs. Ad-GFP HFD group (F). Each assay condition was done in triplicate. Representative images are shown. Paraffin sections of liver samples were subjected to IHC staining, stains without primary antibody were used as negative controls. The liver samples prepared in Fig. 5 were observed by light microscopic examination (x400) G). Each assay condition was done in triplicate. Representative images are shown.