Figure 3

(A) Krox20 expression at the indicated time points following insulin induction.

(B) Insulin-stimulated expression of Krox20 is selectively blocked by a MAPK inhibitor. Inhibitors of various signaling pathways were added to the culture medium 30 min before insulin application. Total RNA samples were harvested 30 min after insulin addition. Error bars represent the standard deviation of three independent experiments.

(C and D) Real-time PCR analysis of the expression of C/EBPβ and KLF5 at the indicated time points following insulin induction (without IBMX/Dex). The expression is normalized to EF2 mRNA expression.

(E) si-Med23 3T3-L1 cells were infected with a retrovirus encoding the Krox20 cDNA and selected with hygromycin. Real-time PCR was used to quantify the Krox20 expression level of the indicated cell lines. The expression is normalized to EF2 mRNA.

(F) Hormone-induced adipogenesis of si-Ctrl, si-Med23, and si-Med23+Krox20 3T3-L1 cells. At day 8 postinduction, cells were stained for lipid droplets with ORO. Scale bar, 100µm.

(G) Real-time PCR analysis of the expression level of adipocyte markers in the indicated 3T3-L1 cell lines at different time points postinduction. The expression is normalized to EF2 mRNA expression.