Fig. 5.

Restoration of MTHFR facilitates chemosensitivity to 5-FU and apoptosis and reduces proliferation of EC cells. Expression vectors containing the MTHFR or containing MTHFR and lncRNA HOTAIR were introduced into TE-1/5-FU cells (oe-MTHFR and oe-MTHFR + oe-lncRNA HOTAIR group). Empty vectors-treated cells were served as negative controls (oe-NC group and oe-MTHFR + oe-NC group). a Western blot analysis was performed to determine the MTHFR expression in TE-1 cells and TE-1/5-FU cells, normalized to GAPDH. b Overexpression efficiency in TE-1/5-FU cells detected using Western blot analysis. c CCK-8 assay was performed to detect TE-1/5-FU cell viability following MTHFR alternations. d CCK-8 assay was performed to measure IC₅₀ value in TE-1/5-FU cells following MTHFR alternations. e EdU assay was performed to measure cell proliferation of TE-1/5-FU cells following MTHFR alternations (× 400). f Flow cytometry was performed to detect apoptosis of TE-1/5-FU cells following MTHFR alternations. g Flow cytometry was performed to detect cell cycle of TE-1/5-FU cells following MTHFR alternations. Values obtained from three independent experiments in triplicate are expressed as mean ± SD and analyzed by ANOVA followed by Tukey’s post hoc test among three or more groups. *p < 0.05 compared with TE-1/5-FU cells treated with either sh-NC or oe-NC group