Extended Data Fig. 3: FTO depletion in prostate, cervical and lung cancers.
a, FTO depletion by shRNA in PC3 prostate cancer cells by RT-qPCR (left). Mean + SD from 3 independent experiments, normalized to ACTB and SDHA. P value calculated by two-tailed paired t-test (P = 0.004). Representative western blot (right) of FTO depletion in PC3 cells (n = 3) with β-actin as loading control. b, Nude mice were injected subcutaneously with PC3 prostate cells and tumors was monitored every two days (n = 6 tumors per group; mean ± SEM, p = 0.0002, two-way ANOVA). c, Real-time chemotactic invasion of RNAi FTO (red) and RNAi control PC3 cells (green). d, FTO depletion by shRNA in Ca Ski cervical cancer cells (left, n = 3, P = 0.009) and H1650 lung cancer cells (right, n = 3, p = 0.0004) by RT-qPCR. Data as mean + SD, normalized to ACTB and SDHA. P values calculated by two-tailed paired t-test. e, Effects of FTO knockdown on tumorsphere-forming capacity in Ca Ski (left, P = 0.02) and H1650 cells (right, p = 0.008). Data from n = 3 technical replicates (mean + SD) within a single experiment, representative of 2 biologically independent experiments provided as Source Data. P values calculated by two-tailed t-test. f, Real-time chemotactic migration of FTO-depleted (red) and control (green) H1650 cells. g-h, Kaplan-Meier curves showing the overall survival of patients with either high-FTO or low-FTO lung tumors (n(high FTO) = 333, n(low FTO) = 326, logrank test, P = 0.0073) (g) or uterine tumors (n(high FTO) = 310, n(low FTO) = 232, logrank test, P = 0.048) (h). Motility data in (c), (f) from n = 3 technical replicates (mean + SD) within a single experiment, representative of 2 and 3 biologically independent experiments, respectively. Independent repeats have been deposited on the Figshare repository (DOI: 10.6084/m9.figshare.14602971).
