Fig. 4
FTO-demethylated m6A promotes gene expression at post-transcriptional levels. a, b qPCR of K562 parental cells transfected with FTO expression or control vectors (a) or nilotinibR cells transfected with FTO shRNA or scramble vectors (b) for 48 h. c qPCR of K562 nilotinibR cells exposed to 25 µM rhein for 48 h. d qPCR of parental and resistant K562 cells treated with actinomycin-D (5 μg/ml) for the indicated time points. The gene expression was normalized to GAPDH. e qPCR of K562 nilotinibR cells transfected with FTO shRNA or scrambled vectors for 48 h followed by 5 μg/ml actinomycin-D treatment for the indicated time points. The gene expression was normalized to GAPDH. f, g K562 parental cells were transfected with YTHDF2 shRNA (f) or nilotinibR cells with YTHDF2 expression (g) vectors for 48 h followed by 5 μg/ml actinomycin-D treatment for the indicated time points. The gene expression was determined by qPCR and normalized to GAPDH. h Parental and resistant K562 cells were treated with 200 nM puromycin for the indicated time points. i Western blotting of K562 parental and resistant cells. j–l K562 nilotinibR cells were transfected with FTO shRNA vectors (j), treated with 25 µM rhein (k) or 50 µM meclofenamic acid (l) for 48 h followed by 200 nM puromycin treatment for the indicated time points. In h and j–l, graphs are the quantification of Western blotting results normalized to β-actin; Data represent three independent experiments; Data are mean ± SD; *p < 0.05, **p < 0.01. See also Figure S3
