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Fig. 3 | Genome Biology

Fig. 3

From: Functional CRISPR screen identifies AP1-associated enhancer regulating FOXF1 to modulate oncogene-induced senescence

Fig. 3

FOXF1 is the target gene regulated by EnhAP1-OIS1. a UCSC screenshot of GRO-seq analysis of BJ-indRASG12V cells. BJ cells were treated with DMSO or 4-OHT for 14 days. Bi-directional transcription is represented by using positive and negative values for expression in the Crick and Watson strands, respectively. The genomic regions of EnhAP1-OIS1 and FOXF1 are enlarged. Note the enhancement in GRO-seq signal for both EnhAP1-OIS1 and FOXF1 in BJ + 4-OHT (brown track) compared to BJ + DMSO (blue track). b mRNA levels of FOXF1 are reduced in sgRNA-AP169 and sgRNA-AP171 targeted cells under 4-OHT treatment. Data shown represent mean (SD), n = 3. *p < 0.05. c BJ-indRASG12V cells transduced with the specified sgRNAs were treated with DMSO or 4-OHT for 14 days; FOXF1, p21, and HRas protein levels were measured by western blot. HSP90 was used as the loading control. The band of FOXF1 is marked with an arrow. ER-HRas indicates the induced version of HRas. d Targeting the FOXF1 and p53 genes caused OIS bypass as measured by β-gal staining. Note the stronger effect of FOXF1ko compared to the effect elicited by targeting EnhAP1-OIS1 (Fig. 2d). Data shown represent mean (SD), n = 4. *p < 0.05. e Targeting FOXF1 and p53 gene resulted in enhanced proliferation as measured by BrdU staining. Data shown represent mean (SD), n = 4. *p < 0.05

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