Figure 2. Effects of decreased NSUN2 and METTL1 expression on 5-FU-treatment and heat stress and 5-FU.

(A) Immunoblot analysis of total proteins lysates and immunoprecipitated NSUN2 and METTL1 proteins from indicated NSUN2-, METTL1-, and double NSUN2/METTL1 knockdowns and control transfectant. As loading control the western blot was probed with an anti-α-tubulin antibody. (B) Time-dependent cell survival in response to heat stress in NSUN2 knockdown cells (open circle), METTL1 knockdown cells (open triangle), NSUN2 and METTL1 knockdown cells (open square), and control vector-transfected cells (closed circle) with the MTT viability assay. There was no significant difference in survival of NSUN2 knockdown cells, METTL-1 knockdown cells, or NSUN2 and METTL1 knockdown cells compared with control vector-transfected cells in all of each duration period. (C) Comparison of the average IC₅₀ values for heat stress between NSUN2 knockdown cells, METTL-1 knockdown cells, or NSUN2 and METTL1 knockdown cells and control vector-transfected cells. NS, not significant. (D) Dose-dependent cell survival in response to 5-FU in NSUN2 knockdown cells (open circle), METTL1 knockdown cells (open triangle), NSUN2 and METTL1 knockdown cells (open square), and control vector-transfected cells (closed circle) with the MTT viability assay. *Significant difference compared with control vector-transfected cells (P<0.05). (E) Comparison of the average IC₅₀ values for 5-FU between NSUN2 knockdown cells, METTL-1 knockdown cells, or NSUN2 and METTL1 knockdown cells and control vector-transfected cells. NS, not significant. *Significant difference compared with control vector-transfected cells (P<0.05).