Fig. 4
From: GRHL2-controlled gene expression networks in luminal breast cancer
EHF represents a direct GRHL2 regulated gene. a Bru-seq reads for EHF at indicated time points after to GRHL2 deletion. Track colors: green, sgCTR; red, sgGRHL2(1); blue, sgGRHL2(2). Positive y-axis indicates the plus-strand signal of RNA synthesis from left to right and the negative y-axis represents the minus-strand signal of RNA synthesis from right to left. b Line graph depicting the log2 AFC of EHF transcription in sgGRHL2(1) and sgGRHL2(2) cells. c ChIP-qPCR showing enrichment of GRHL2 binding sites in EHF promoter region but not in the control GAPDH gene. Graph represents the efficiency of indicated genomic DNA co-precipitation with anti-GRHL2 Ab (black bars) or IgG control Ab (grey bars). Signals for IgG control and GRHL2 antibody pulldown samples are normalized to input DNA and are presented as % input with SEM from 3 technical replicates. Data are statistically analyzed by t-test and *indicates p < 0.05. d EHF mRNA expression in a panel of 52 human breast cancer cell lines covering luminal-, basal A-, and basal B subtypes extracted from RNA-seq data. Data is statistically analyzed by t-test and *indicates p < 0.05. e qRT-PCR analysis of expression level of EHF mRNA after 4 days of doxycycline treatment of MCF7 cells transduced with dox-inducible Cas9 and sgCTR or sgGRHL2 constructs, in combination with ectopic expression of EHF or empty vector (EV) plasmids. Data are presented as mean ± SEM from three technical replicates. Data are statistically analyzed by t-test. *Indicates p < 0.05. f Graph showing results from SRB assay after 4 days doxycycline-treatment as in (e) and subsequent culture for the indicated time periods