Fig. 5
FTO promotes TKI resistance. a, b Parental cells were transiently transfected with FTO expression (a) or resistant cells with FTO shRNA (b) vectors and subjected to colony assays in medium free from nilotinib. c CCK-8 assays for parental cells transfected with FTO expression or control vectors for 24 h followed by nilotinib or PKC412 treatment for another 48 h. d Left, Western blotting or dotblotting of K562 parental cells transfected with METTL3 shRNA or scramble vectors for 48 h. Data represent three independent experiments; right, CCK-8 assays for K562 parental cells transfected with METTL3 shRNA or scramble vectors for 24 h followed by nilotinib treatment for additional 48 h. e CCK-8 assays for resistant cells transfected with FTO shRNA or scramble vectors for 24 h followed by nilotinib or PKC412 treatment for another 48 h. f The parental cells were transfected with FTO shRNA or scramble vectors for 24 h and subjected to colony assays in drug-free medium (left) or CCK-8 assays (right) after nilotinib treatment for another 48 h. g External view of tumors, and graphs show the quantification of tumor weight or tumor volume. h Left, representative images of IHC and H&E staining. Window stands for the enlarged cells; right, graph is the quantification of IHC staining. i Left, representative images of m6A dotblotting from tumors; right, graph is the quantification of blot intensities. j IP by anti-m6A antibody was performed in mRNA extracted from tumors. The eluted mRNA was subjected to cDNA synthesis followed by qPCR for gene expression. k qPCR for gene expression in mRNA extracted from tumors. Data in colony assays represent two independent experiments with four duplicates; Data in CCK-8 assays represent two independent experiments with 8 repeats; In j, k, n = 6 tumors/group; Data are mean ± SD; *p < 0.05, **p < 0.01. See also Figures S4, S5 and S6
