Fig. 2
Inhibition of ALKBH5 impairs keratinocyte migration in vitro. a, b Decreased ALKBH5 expression was confirmed in HaCaT cells after siRNA transfection by qRT‒PCR (a) and WB (b). The experiments were performed in triplicate. One-way ANOVA, ***P < 0.001. The full-length blots are presented in Additional file 8: Fig. S1. c Global m6A levels of mRNA extracted from ALKBH5-knockdown or control HaCaT cells after siRNA transfection were determined by m6A DB assays. Total RNA was determined by methylene blue staining (as loading control). d Cell migration of ALKBH5-knockdown or control HaCaT cells was analyzed by Transwell assay. Scale bar: 25 μm. e Statistical analysis of the Transwell assay. All of the experiments were performed in triplicate, and five random fields were included in the analysis. The percentage of migrated cell areas is shown as the mean ± SD. One-way ANOVA, ***P < 0.001. f Representative dynamic cell velocity mapping of ALKBH5-knockdown or control HaCaT cells at 0 h, 2 h, 16 h, and 32 h after scratching using cell tracking imaging. u velocity in the x direction, v velocity in the y direction. The velocity magnitude represents the resultant velocity of u and v. The direction and magnitude of the velocity are indicated with black arrows and color density, respectively. FC phase‒contrast image. Scale bar: 250 μm. g Statistical analysis of the velocity magnitude of HaCaT cells in the ALKBH5 knockdown and control groups at different time points. h Statistical analysis of the healing rate of scratched wounds. All of the experiments were performed in triplicate. i The long-term migratory ability of HaCaT cells was evaluated with wound scratching assays. Scale bar: 100 μm. j Statistical analysis of wound healing assays. All of the experiments were performed in triplicate, and 3 random fields were included at each time point for analysis. One-way ANOVA, ***P < 0.001