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. 2014 Sep 18;10(9):e1004639. doi: 10.1371/journal.pgen.1004639

Figure 6. Effects of co-expression of NSUN2 and METTL1 on the cytotoxic effects of heat stress and 5-FU in NSUN2 and METTL1 knockdown cells.

Figure 6

(A) Immunoblot analysis of proteins from NSUN2 and METTL1 knockdown cells co-expressing Xpress-tagged wild type NSUN2 and Xpress-tagged wild type METTL1 (lane WT/WT), NSUN2 and METTL1 knockdown cells co-expressing Xpress-tagged phosphorylation-mimic NSUN2 and Xpress-tagged phosphorylation-mimic METTL1 (lane SE/SE), NSUN2 and METTL1 knockdown cells co-expressing Xpress-tagged dephosphorylation-mimic NSUN2 and Xpress-tagged dephosphorylation-mimic METTL1 (lane SA/SA), and NSUN2 and METTL1 knockdown cells transfected with control vector (lane empty) with anti-Xpress and anti-α-tubulin antibodies. Five independent clones that co-expressed Xpress-NSUN2 and Xpress-METTL1 or were transfected with the empty vector, originating from UTR-targeting shRNA-mediated NSUN2 and METTL1 knockdown cells, were pooled and used as a stable transfectant. (B) Dose-dependent cell survival in response to 5-FU in NSUN2 and METTL1 knockdown cells co-expressing Xpress-tagged wild type NSUN2 and Xpress-tagged wild type METTL1 (open circle, DKD + WT/WT), NSUN2 and METTL1 knockdown cells co-expressing Xpress-tagged phosphorylation-mimic NSUN2 and Xpress-tagged phosphorylation-mimic METTL1 (open triangle, DKD + SE/SE), NSUN2 and METTL1 knockdown cells co-expressing Xpress-tagged dephosphorylation-mimic NSUN2 and Xpress-tagged dephosphorylation-mimic METTL1 (open square, DKD + SA/SA), and NSUN2 and METTL1 knockdown cells transfected with control vector (closed circle, DKD + empty) with the MTT viability assay. (C) Comparison of the average IC50 values for 5-FU between NSUN2 and METTL1 knockdown cells co-expressing NSUN2 and METTL1 and control vector-transfected cells. NS, not significant. *Significant difference compared with control vector-transfected cells (P<0.05).