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. 2011 Dec 13;31(4):972–985. doi: 10.1038/emboj.2011.451

Figure 7.

Figure 7

E2F3 and HELLS regulate many common targets, most notably MLL1. (A) Confirmatory ChIP analyses of HELLS target genes in DU145 prostate carcinoma cells. The enrichment of E2F3 and HELLS was assessed using PDS5A, KRAS, MLL1, HSPD1 and RNF146 primers spanning the genomic regions around the TSS. CCNA2 is an E2F-dependent promoter and U2 served as a negative promoter (Ctrl) and IgG served as antibody control. (B) qRT–PCR (qPCR) analysis depicting the relative expression of representative HELLS target genes in human prostate carcinoma cells (PC3) after lentiviral depletion of E2F3 or HELLS using two independent sequences each. The knockdown was verified by qPCR analysis of E2F3 and HELLS. The transcriptional regulation of PDS5A, KRAS, MLL1, HSPD1, RNF146, SLC44A1, ZNF76 (M. musculus Zfp523), XPO4, RALGPS2 and NEK6 was analysed. GAPDH served as a control. (C) MLL1 was analysed by qRT–PCR in synchronized PC3 cells, which were serum-arrested and harvested at 0 or 18 h after serum addition. CDC6 is a S-phase-specific control. (D) Effects of depleting HELLS in PC3 cells determined by western blots for MLL1, HELLS and TUBULIN, using two independent hairpins (shHELLS S1 or S2) or control infections (shCtrl). Figure source data can be found in Supplementary data.