Fig. 4

MGAT4EP is predominantly localized in the nucleus and interacts with transcription factor FOXA1. A The RNA level of MGAT4EP in nuclear and the cytoplasmic fraction of MCF7 and T47D cells was measured by qRT-PCR. MALAT1 RNA and GAPDH mRNA was used a positive control for nuclear and cytoplasmic fraction, respectively. B The proteins retrieved by RNA pull-down with MGAT4EP RNA and negative control antisense RNA (AS) were visualized by silver staining and subject to mass spectrometry (MS) analysis. C RNA pull-down coupled with western blot validated the interaction between MGAT4EP and FOXA1 that was identified from MS analysis. SP1 that was not found in MS analysis was used as a negative control. D RNA pull-down of the antisense, full-length, and serial deletion mutants of MGAT4EP RNA followed by anti-FOXA1/anti-SP1 western blotting. The four serial deletion mutants of MGAT4EP RNA were generated by deleting 1–700, 700–1400, 1400–2100, or 2100–2819 bps, respectively. E RIP-qPCR analysis with anti-FOXA1 or anti-IgG antibody validated the association of FOXA1 with MGAT4EP RNA, where MALAT1 and GAPDH RNA were used as negative controls. 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)