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. 2021 Apr 3;11(12):5926–5938. doi: 10.7150/thno.56490

Figure 3.

Figure 3

Three E2-box-like sequences in the promoter region of PFKM are not required for its transactivation by ZEB1. (A) MHCC-97H cell line was knocked down and further re-expressed for ZEB1. Then relative mRNA levels of ZEB1 and PFKM were determined using RT-qPCR. (B) HEK-293T cells were transfected with PFKM-Luc and increasing doses of ZEB1. 24 h post-transfection, luciferase activity was determined. The data are presented as means±SD of three independent experiments. Statistical analysis was performed by one-way ANOVA. (C) Schematic diagram showing wildtype PFKM promoter and its various mutants with three E2-box-like sequences mutated alone or in combinations. For each E2-box-like sequence, CACCT(G) was mutated to CAAAT(G). (D) Luciferase reporter vectors carrying either wildtype PFKM promoter or any of its mutants was transfected into HEK-293T cells together with ZEB1. After 24 h of transfection, relative luciferase activity was determined and normalized to cells transfected with wildtype PFKM promoter alone (column 1). The data in (A) and (D) are shown as means±SD of three independent experiments (***P<0.001, N.S.: P ≥ 0.05, unpaired Student's t test).