Fig. 6
NSUN2-mediated mRNA m5C methylation promotes HGH1 expression (A) RT-qPCR assays were used to measure the expression of HGH1 in BC cell lines after NSUN2 knockdown, and (B) Western blot analysis was used to confirm the protein level. (C) and (D) RT-qPCR and Western blot assays were used to assess alterations in HGH1 expression after overexpression of NSUN2-WT or NSUN2-DM. (E) MeRIP-qPCR was used to measure the level of m5C on HGH1 mRNA after NSUN2 knockout in BC cells. (F) Changes in the response to actinomycin D-mediated RNA synthesis inhibition and mRNA transcription interference were detected after NSUN2 knockdown. (G) and (H) CCK-8 experiments confirmed the influence of NSUN2 and HGH1 on the proliferation of BC cells and their downstream relationship. (I) A colony formation assay was used to determine the effect of restoring the expression of the downstream gene HGH1 in the NSUN2-knockdown cell line on the expansion ability of BC cells. ns. no significant; Data are mean ± SD of three independent experiments. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001 by t test