Fig. 5.

PD-1 (PDCD1), CXCR4, and SOX10 are critical target genes of FTO in melanoma cells. a, b Immunoblot analysis of PD-1, p-p70S6K, p70S6K, CXCR4, SOX10, FTO, and β-actin in Mel624 stable cells with or without knockdown (a) and forced overexpression (b) of FTO. c Immunoblot analysis of PD-1, p-p70S6K, p70S6K, FTO, and β-actin in Mel624 with or without FTO knockdown, and/or PD-1 (PDCD1) overexpression. d Cell proliferation assay in cells as in c. e Cell migration assay in cells as in c. f, g Cell proliferation (f) and migration analysis (g) in Mel624 with or without FTO knockdown, and/or CXCR4 overexpression. h, i Cell proliferation (h) and migration analysis (i) in Mel624 with or without FTO knockdown, and/or SOX10 overexpression. j–n qPCR analysis of the expression of PD-1 (PDCD1) (j), CXCR4 (k), SOX10 (l), CTSV (m), and NOP16 (n) in Mel624 cells with or without FTO knockdown, in combination with siRNA knockdown of both METTL3 and METTL14. o Immunoblot analysis of PD-1, CXCR4, SOX10, FTO, METTL3, METTL14, and GAPDH in cells as in j–n. Data are shown as mean ± S.D. (d–i), or mean ± S.E. (j–n) (n ≥ 3). *P < 0.05; **P < 0.01; ***P < 0.001; Student’s t-test