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. 2021 May 27;12(6):550. doi: 10.1038/s41419-021-03797-3

Fig. 6. Transcription factor E2F1 directly binds to the promoter of SEC61G and regulates the expression of SEC61G in breast cancer cells.

Fig. 6

A The putative binding motif of E2F1 was predicted by using the JASPAR database (http://jaspar.generg.net) and Matrix profile. B The diagram showed the putative WT TEAD binding elements (TBE) located in the promoter region of the SEC61G gene. Luciferase reporter vectors containing WT TBE1/TBE2 (P1), mutated TBE1 (P1-M1), or mutated TBE2 (P1-M2) were constructed. C Luciferase reporter vectors containing WT or mutated TBE sequences were co-transfected with control or E2F1 overexpression vector into HEK293 cells. The relative luciferase activity was analyzed 48 h later. D HEK293 cells were transfected with siRNA-NC or siRNA-E2F1 and then RNA immunoprecipitation Chip (RIP) assay was performed using control IgG or SEC61G antibody. The SEC61G enrichment was analyzed by qPCR. E–G MCF-7 or MDA-MB-231 cells were transfected with siRNA-NC, siRNA-E2F1, empty vector, E2F1 overexpress vector (OE), or left untreated (Blank). E The protein expression of E2F1 and SEC61G was analyzed by western blot. GAPDH was used as a loading control. F, G The relative expression of SEC61G mRNA in MCF-7 or MDA-MB-231 cells was analyzed by qPCR. *P < 0.05, **P < 0.01.