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Fig. 5 | Genome Biology

Fig. 5

From: Systematic functional interrogation of human pseudogenes using CRISPRi

Fig. 5

MGAT4EP upregulates the expression of FOXM1, a FOXA1 target, and enhances FOXA1 binding to its promoter. A Schema depicting the workflow of identifying potential protein-coding gene targets that were co-regulated by MGAT4EP and FOXA1 and were important for mediating their tumor-promoting function in luminal A breast cancer. B QRT-PCR analysis of FOXM1 mRNA expression and C western blot for measuring FOXM1 protein expression in MCF7 and T47D cells that were treated with negative control non-targeting siRNA (si-NC) or MGAT4EP-targeting siRNAs. D qRT-PCR analysis of FOXM1 mRNA expression and E western blot for measuring FOXM1 protein expression in MCF7 cells and T47D cells that were treated with si-NC or FOXA1-targeting siRNAs. F The signal track of FOXA1 ChIP-seq and the corresponding input in MCF7 and T47D cells. The identified ChIP-seq peaks were drawn as horizontal lines above the signal track. G ChIP-qPCR analysis was performed with anti-FOXA1 or anti-IgG antibody in MCF7 and T47D cells to confirm the enrichment of DNA fragments covering the FOXA1 ChIP-seq peak in the FOXM1 promoter. The effect of si-NC or MGAT4EP-targeting siRNAs on the binding of FOXA1 to the same region was assessed by ChIP-qPCR. H Western blot for measuring FOXM1 protein expression in MCF7 cells and T47D cells that were transduced with negative control non-targeting shRNA (sh-NC) or FOXM1-targeting shRNAs. I The growth of MCF7 and T47D cells transduced with sh-NC or FOXM1-targeting shRNAs was monitored (OD450 absorbance for WST-8 formazen) every 24 h with CCK-8 assay for 96 h. All data are shown as mean ± SD, n = 3. The Student’s t test was used to assess the statistical significance of difference in mean between two experimental groups (*p < 0.05; **p < 0.01; ns: not significant, p ≥ 0.05)

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