Fig. 5: Essential role of PAK4-mediated phosphorylation of CDK2 in adipogenesis.
From: PAK4 phosphorylates cyclin-dependent kinase 2 to promote the G1/S transition during adipogenesis
a 3T3-L1 preadipocytes were pretreated with 30 nM ND201651 before MDI treatment. Co-IP was performed on day 1 to assess CDK2 phosphorylation and its interaction with C/EBPβ. b–e 3T3-L1 preadipocytes were transfected with either phosphodeficient CDK2 (CDK2S106A, SA) or phosphomimetic CDK2 (CDK2S106D, SD), then induced to differentiate with MDI treatment. After 24 h, C/EBPβ phosphorylation (b) and C/EBP-transcriptional activity (c, n = 3) were analyzed. d Lipid accumulation was assessed after 8 days of differentiation using Oil Red O staining and BODIPY immunostaining. e CDK2 ubiquitination was analyzed in CDK2 immunoprecipitates from cell lysates collected 24 h after the initiation of differentiation following transfection. f Nuclear translocation of CDK2 was examined by western blotting after 24 h of differentiation, with or without 30 nM ND201651 treatment. g 3T3-L1 preadipocytes were treated with 30 nM ND201651, followed by differentiation induction with MDI for 24 h. C/EBP-luciferase activity was then measured (n = 3). Veh, vehicle; RLU, relative luciferase unit; NE, nuclear extract; CE, cytosolic extract.. Values are mean ± s.d. *P < 0.05, **P < 0.01.
