Fig. 6

HINT2 protein expression was regulated by m⁶A modification. a Reduction in m⁶A modification in specific regions of HINT2 transcripts upon METTL3 knockdown, as assessed by gene-specific m⁶A-RIP-qPCR assays, in PIG1, OCM1 and CRMM1 cells. The value obtained for the control group was set to 1. Error bars indicate the mean ± SEM, n = 3, *p < 0.05, **p < 0.01. b Reduction in m⁶A modification in specific regions of HINT2 transcripts upon ALKBH5 knockdown, as assessed by gene-specific m⁶A-RIP-qPCR assays, in OCM1 and CRMM1 cells. The value obtained for the control group was set to 1. Error bars indicate the mean ± SEM, n = 3, *p < 0.05, **p < 0.01. c, d Western blot (Left) and real-time PCR (Right) showing HINT2 expression in PIG1 cells with or without METTL3 knockdown (c) and in OCM1 and CRMM1 cells with or without ALKBH5 knockdown (d). The HINT2 signal was quantified and normalized to that of β-actin. Error bars indicate the mean ± SEM, n = 3. e HINT2 expression in OCM1 and CRMM1 cells with or without ALKBH5 knockdown and with or without HINT2 knockdown. f A colony formation assay was performed to assess the tumor growth of OCM1 and CRMM1 cells with or without ALKBH5 knockdown and with or without HINT2 knockdown. g Statistical analysis of the colony formation assay performed using OCM1 and CRMM1 cells with or without ALKBH5 knockdown and with or without HINT2 knockdown. The colony number in the control group was set to 1. All of the experiments were performed in triplicate, and relative colony formation rates are shown as the mean ± SEM. n = 3, **p < 0.01, ***p < 0.001. h Proliferation of OCM1 and CRMM1 cells with or without ALKBH5 knockdown and with or without HINT2 knockdown, as determined by CCK8 assays. n = 3, ∗∗p < 0.01