Extended Data Fig. 5: YTHDF2 deficiency improves macrophage anti-tumoral polarization by targeting IFN-γ-STAT1 signaling.
a, Ingenuity pathway analysis showing the top four pathways enriched in Ythdf2cKO anti-tumoral macrophages compared to Ythdf2f/f anti-tumoral macrophages. P values were determined by hypergeometric test and adjusted for multiple testing using the Benjamini–Hochberg method. b, qPCR showing the expression of IFN-γ response genes in BMDMs from Ythdf2f/f and Ythdf2cKO mice that were treated with IFN-γ (100 ng/ml) for 6 h (n = 3). c, qPCR showing the expression of Stat1 in CD11b+F4/80+ TAMs isolated from tumor tissues of B16-OVA tumor-bearing Ythdf2f/f and Ythdf2cKO on day 14 post B16-OVA tumor inoculation (n = 4). d, Immunoblotting showing the expression of phosphor (p)-STAT1 and STAT1 in CD11b+F4/80+ TAMs isolated from tumor tissues of B16-OVA tumor-bearing Ythdf2f/f and Ythdf2cKO mice on day 14 post-B16-OVA tumor inoculation. e, Immunoblotting showing the expression of STAT1 in BMDMs transfected with Stat1 siRNA or control siRNA. f, Immunoblotting showing the expression of STAT1 in BMDMs nucleofected with Stat1 gRNA or control gRNA. g, Representative histogram of the expression of IFNGR1 in BMDMs nucleofected with Ifngr1 gRNA or control gRNA. h, B16-OVA tumor growth on day 18 post-tumor inoculation in Rag1−/− mice s.c. implanted with BMDMs from Ythdf2f/f and Ythdf2cKO mice together with i.v. injection of CD8+ T cells from IFN-γ−/− mice (n = 4). Data are shown as mean ± SD and were analyzed with an unpaired two-tailed t-test (b, c) or two-way ANOVA with mixed-effects model and adjusted by the Holm-Šídák post-test (h). The numbers below a lane indicate relative expression levels of target proteins normalized to the expression of β-actin. Data in b-h are representative of at least two independent experiments. NS, not significant.
