Fig 1: NSUN2 stabilizes KRT13 mRNA by recruiting YBX1. (A) Effect of NSUN2 on the stability of KRT13 mRNA. CaSki cells transfected with two NSUN2-targeting shRNAs were treated with 10 µg/ml RNA synthesis inhibitor Actinomycin D (Actd) for indicated time. Level of KRT13 mRNA was analyzed by RT-qPCR. (B, C) Effect of YBX1 (B) and ALYREF (C) knockdown on KRT13 mRNA and protein level in CaSki cells as assessed by RT-qPCR and Western blot. (D, E) Effect of YBX1 (D) and ALYREF (E) overexpression on KRT13 mRNA and protein level in CaSki cells as assessed by RT-qPCR and Western blot. (F) RIP-qPCR analysis of YBX1-KRT13 interaction. IgG was used as a negative control. (G, H) Effect of YBX1 on m5C modification occurred on KRT13 mRNA. YBX1 was silenced (G) or overexpressed (H) in CaSki cells, and the m5C modification on KRT13 mRNA was measured by m5C-MeRIP-qPCR. NAPRT1 was used as a positive control. KRT13-Ctrl was used as a negative control. (I) A working model of the mechanism by which NSUN2 promotes cervical cancer migration and invasion. (Data shown are mean ± SEM, n = 3; Student’s unpaired t-test was used for statistical analysis, **p < 0.01, ***p < 0.001).