Fig. 4.

KAP1 interacts with ZFP30 and promotes adipogenesis. a Immunoprecipitation in HEK293T cells showing that ZFP30 interacts with KAP1. HEK293T cells transfected with the indicated constructs were subjected to immunoprecipitation with the HA antibody and subsequently probed with the listed antibodies. b Immunoprecipitation in 3T3-L1 cells showing that ZFP30 interacts with endogenous KAP1 in adipogenic conditions. 3T3-L1 expressing the indicated HA-tagged proteins were used for immunoprecipitating HA and subsequent western blotting with the listed antibodies. NS, non-specific band. c Immunoprecipitation in HEK293T cells showing that the KRAB domain of ZFP30 is essential for interaction with KAP1. ΔKRAB is a KRAB domain deletion mutant and In24aa is a mutant with a 24 amino acid insertion in the KRAB domain derived from the Zfp30 c15 allele 2. d Effect of knocking down Kap1 and of rescuing Kap1 expression on IBA adipogenic differentiation as assessed by lipid accumulation (ORO staining). The rescue of Kap1 expression was achieved by introducing an shRNA-resistant allele. The expression of endogenous and exogenous Kap1 (endo-Kap1 and exo-Kap1, respectively), as well as adipogenic marker genes was assessed by qPCR. n = 3 biologically independent experiments. **p < 0.01, *p < 0.05, t test, compared with shControl. e Effect of knocking out Kap1 on primary MEF adipogenic gene expression as assessed by qPCR. Lentivirus encoding Cre recombinase or control vector were introduced to Kap1^loxp/loxp MEF cells to generate Kap1 KO and control cells. KAP1 protein levels are shown on the right. n = 3 biologically independent experiments. **p < 0.01, *p < 0.05, t test. All panels: error bars—SD. Source data are provided as a Source Data file