Figures & data
Figure 1. Mstn knockdown reduced the formation of lipid droplets and affected the expression of adipocyte-specific genes. (a, b) MSTN mRNA and protein levels after Mstn knockdown in C2C12 myoblasts. (c) Oil Red O staining of lipid droplets after induction of adipogenic differentiation in C2C12 myoblasts. Left panel: lipid droplets formed from Mstn RNAi C2C12 myoblasts; right panel: control. Lipid droplets were marked with white arrows. (d) Absorbance of lipid droplets at 520 nm. (e) Effects of Mstn knockdown on the expression of the anti-adipogenic factor Gata2 and adipocyte-specific genes. *p < 0.05, **p < 0.01, ***p < 0.001; t-tests were used to generate p values. shRNA C: control shRNA.
Figure 2. Mstn knockdown increased the H3K27me3 modification on adipocyte-specific genes by reducing the expression of the histone demethylase Jmjd3. (a) Schematic diagram of the detected binding region of H3K27me3 modification. The detected binding region was located on the promoter of adipocyte-specific genes. (b) The H3K27me3 modification on adipocyte-specific genes was detected by ChIP-qPCR. (c, d) JMJD3 mRNA and protein levels after Mstn knockdown. *p < 0.05, **p < 0.01, ***p < 0.001; t-tests were used to generate p values. Con: control.
Figure 3. Mstn regulated the expression of Jmjd3 via SMAD2/SMAD3. (a) Binding region of SMAD2/SMAD3 with the promoter of Jmjd3. (b) The promoter of Jmjd3 combined with SMAD2/SMAD3 detected by ChIP-qPCR. The detected binding region was from −1374 bp to −1236 bp on the promoter of Jmjd3.
Figure 4. Overexpression of Jmjd3 erased the H3K27me3 markers and promoted the expression of adipocyte-specific genes. (a, b) JMJD3 mRNA and protein levels after Jmjd3 overexpression in C2C12 myoblasts. (c) H3K27me3 modification on adipocyte-specific genes after overexpression of Jmjd3. (d) Expression of adipocyte-specific genes after overexpression of Jmjd3. *p < 0.05, **p < 0.01, ***p < 0.001; t-tests were used to generate p values. + Jmjd3: overexpression of Jmjd3.
