Figures & data
Figure 1. Gene expression during adipose differentiation of 3T3-F442A cells. (A) Adipose conversion shown by lipid staining with Oil Red O. Staurosporine (St) at low concentrations [10–12 nM] induced adipose differentiation in 3T3-F442A cells and this effect was augmented by Dexamethasone (Dex) at [250 nM]. Dex alone did not induce adipose differentiation; (N-Ad, non-adipogenic medium). (B) mRNA expression of adipogenic genes Cebpb, Pparg2, Cepba, Klf4 and Klf5 during adipose differentiation of 3T3-F442A cells treated with St-Dex. (C) Expression of Klf4 at 2h, Cebpb at 4h,and Klf5 at 3h, which are the times of their highest expression.
![Figure 1. Gene expression during adipose differentiation of 3T3-F442A cells. (A) Adipose conversion shown by lipid staining with Oil Red O. Staurosporine (St) at low concentrations [10–12 nM] induced adipose differentiation in 3T3-F442A cells and this effect was augmented by Dexamethasone (Dex) at [250 nM]. Dex alone did not induce adipose differentiation; (N-Ad, non-adipogenic medium). (B) mRNA expression of adipogenic genes Cebpb, Pparg2, Cepba, Klf4 and Klf5 during adipose differentiation of 3T3-F442A cells treated with St-Dex. (C) Expression of Klf4 at 2h, Cebpb at 4h,and Klf5 at 3h, which are the times of their highest expression.](/cms/asset/505cb489-31bd-4a3e-8782-96c70a766c36/kadi_a_1007823_f0001_oc.gif)
Figure 2. Forced expression of Klf4 does not increase Cebpb and Klf5 expression. (A) Adipose conversion shown by lipid staining with Oil Red O. (B) Expression of the adipogenic genes Klf4, Cebpb, Klf5 and Pparg2 in cultures transfected with pCMVSport6 empty vector (pS-empty) and pCMVSport6Klf4 (pS-Klf4) in cells cultured in non-adipogenic conditions. (C) Representative western blot showing the amounts of KLF4 and using GDI protein as loading control.
Figure 3. GSK3β activity and retinoic acid affect gene expression during adipose differentiation. (A) Adipose conversion shown by lipid staining with Oil Red O. GSK3β inhibitor (SB415286) at [100μM] or retinoic acid at [10μM]. (B and C) Expression of Pparg2, Cebpa, Klf4, Cebpb and Klf5 in 3T3F442A St-Dex cultures treated with SB415286 (black) or retinoic acid (blue). (D) Adipose conversion shown by lipid staining with Oil Red O in cultures treated with Cycloheximide (CHX) at [15μM]. (E) Expression of Klf4, Cebpb, and Klf5 in cultures treated with or without CHX for 8 h in St-Dex induced cultures.
![Figure 3. GSK3β activity and retinoic acid affect gene expression during adipose differentiation. (A) Adipose conversion shown by lipid staining with Oil Red O. GSK3β inhibitor (SB415286) at [100μM] or retinoic acid at [10μM]. (B and C) Expression of Pparg2, Cebpa, Klf4, Cebpb and Klf5 in 3T3F442A St-Dex cultures treated with SB415286 (black) or retinoic acid (blue). (D) Adipose conversion shown by lipid staining with Oil Red O in cultures treated with Cycloheximide (CHX) at [15μM]. (E) Expression of Klf4, Cebpb, and Klf5 in cultures treated with or without CHX for 8 h in St-Dex induced cultures.](/cms/asset/d7811899-d78d-428c-8cfd-ea4b0753d53a/kadi_a_1007823_f0003_oc.gif)
Figure 4. Transcriptional cascade showing Klf4 and Klf5 expression during adipogenesis. The adipogenic program involves a transcriptional cascade with early participation of KLF4 and KLF5, which depends on GSK3β activity. Forced expression of Klf4 showed that Cebpb expression was not dependent on KLF4. All-trans retinoic acid impairs the adipogenic program after Cebpb expression and up-stream of Klf4 and Klf5 expression. Solid line means direct relationship and dashed line means an indirect or multistep involved relationship. This schematic was constructed from cited references 1 and 3.
