Fig. 5.

ZFP30 activates Pparg2 expression through KAP1. a Number of KAP1 ChIP-seq peaks in 3T3-L1 cells at days 0 and 2 upon adipogenesis. Two biological replicates (R1 and R2) were performed. b The overlap between KAP1 ChIP-seq peaks at days 0 and 2. c The fraction of distinct categories of peaks (union of all KAP1 – KAP1.u, KAP1 present in at least two samples – KAP1.i, ZFP30, ZEB1, or p300) proximal to a gene encoding a KZFP. d KAP1, ZFP30-HA, RNA PolII (POL2), p300/CBP, H3K4me1, and H3K27ac (K27Ac) ChIP-seq enrichments at the Pparg2 locus in 3T3-L1 cells (upper panel, gray rectangle) or brown pre-adipocytes (lower panel, brown rectangle) at days 0 and 2 of adipogenic differentiation. e ChIP qPCR probing IgG or KAP1 in 3T3-L1 cells upon Zfp30 KD or shRNA control at days 0 and 2. The KAP1-bound Pparg and Zfp810 loci as well as a negative control were tested. n = 3 biologically independent experiments. **p < 0.01, t test. f Luciferase assay showing that KAP1 is essential for the identified Pparg2 enhancer activity. Transcriptional activity as measured by luciferase assays (Firefly/Renilla) for Pparg2-P and Pparg2-PdM (see Fig. 3g) at days 2 of adipogenic differentiation upon Kap1 KD and rescue, as shown in Fig. 4d. n = 3 biologically independent experiments. **p < 0.01, t test. g, h Rescue of the differentiation defect in Kap1 KD and Zfp30 KO IBA cells, respectively, by ectopic expression of Pparg2. Pparg2 expression can be induced by Doxycycline (Dox). Differentiation was assessed by lipid accumulation (ORO staining) while the protein levels were measured by western blotting. Note that there is already leaky expression of Pparg2 when Dox is absent. All panels: error bars–SD. Source data are provided as a Source Data file