Fig. 7
From: m6A mRNA demethylase FTO regulates melanoma tumorigenicity and response to anti-PD-1 blockade
Metabolic stress induces FTO expression through NF-kB and autophagy. a qPCR analysis of FTO and PD-1 (PDCD1) mRNA levels in Mel624 cells cultured with control medium (10% FBS DMEM), 0.2% FBS DMEM, serum-free DMEM, Hanks’ balanced salt solution containing calcium and magnesium (HBSS), or a combination of DMEM and HBSS. b Immunoblot analysis of FTO, PD-1, p62, LC3-I/II (short and long exposure), and GAPDH in Mel624 cells treated as in a. c m6A dot blot assays using total RNA of Mel624 cultured with control medium or HBSS. Methylene blue staining was used as a loading control. d qPCR analysis of FTO and PD-1 (PDCD1) mRNA levels in Mel624 cells with or without knockdown of ATG5 or ATG7 and with or without metabolic stress. e Luciferase reporter analysis of NF-κB response element in Mel624 cells as in d. f qPCR analysis of FTO and PD-1 (PDCD1) mRNA levels in Mel624 cells with or without FTO knockdown treated with or without metabolic stress. g qPCR analysis of FTO and PD-1 (PDCD1) mRNA levels in Mel624 cells with or without siRNA knockdown of RELA treated as in e. Data are shown as mean ± S.E. (a, d, f, g), or mean ± S.D. (e) (n ≥ 3).*P < 0.05; **P < 0.01; ***P < 0.001; Student’s t-test
