Figure 2

Figure 2 FTO promotes endothelial cell cycle progression and tip cell formation to facilitate angiogenesis in vitro.

(A) GSEA plot of pathways enriched in HUVECs transduced with L-FTO is presented. (B, C) Diagram (B) and heatmap (C) show differential expression of genes involved in cell cycle checkpoints in HUVECs transduced with L-EV or L-FTO. (D, E) Flow cytometric analyses on untreated HUVECs, HUVECs transduced with L-EV/L-FTO (D), or transfected with scramble siRNA/FTO-siRNA (E). n = 3 per group. (F, G) EdU assay on untreated HUVECs, HUVECs transduced with indicated lentivirus (F) or transfected with distinct siRNAs (G). Cell nuclei are counterstained with DAPI. n = 3 per group. Scale bar: 60 µm. (H, I) Immunoblotting of cleaved caspase-3 in untreated HUVECs, HUVECs transduced with L-EV/L-FTO (H), or scramble siRNA/FTO-siRNA (I). GAPDH is used as an internal control. n = 3 per group. (J, K). mRNA levels of tip cell markers (CXCR4, FSCN1, APLN, ESM1 and PLAUR) detected by qPCR in HUVECs transduced with indicated lentivirus (J) or transfected with distinct siRNAs (K). n = 3 per group. (L, M) Transwell migration assay on untreated HUVECs or HUVECs transduced with indicated lentivirus (L) or transfected with distinct siRNAs (M). n = 3 per group. Scale bar: 50 µm. (N, O) Scratch test on untreated HUVECs or HUVECs transduced with indicated lentivirus (N) or transfected with distinct siRNAs (O). n = 3 per group. Scale bar: 100 µm. (P, Q) RTCA system demonstrates migration rates of HUVECs transduced with L-EV/L-FTO (P), or transfected with scramble siRNA/FTO-siRNA (Q). n = 3 per group. (R, S) Tube formation assay on untreated HUVECs or HUVECs transduced with L-EV/L-FTO (R), or transfected with scramble siRNA/FTO-siRNA (S). n = 3 per group. Scale bar: 100 µm. Data information: Data represent different numbers (n) of biological replicates. Data are shown as mean ± SEM. One-way ANOVA followed by Bonferroni’s test is used in (D–I, L–O, R, S). Two-tailed Student’s t test is used in (J, K, P, Q). NS: not significant (p > 0.05); *p < 0.05; **p < 0.01; and ***p < 0.001. Source data are available online for this figure.