Fig. 5

PF-502 induces cell-cycle arrest and inhibits endothelial (pHUVEC) proliferation in vivo and in vitro (A) Cell viability of CT26 and pHUVECs was assessed after treated with PF-502(0?10 ??) for 24 h. (B) The ratio of EdU-positive endothelial cells in pHUVECs with or without PF-502 (0.15 ?M for 24 h) treatment. The EdU-positive endothelial cells were analyzed by ImageJ. Scale bars: 50 ?m. Data are expressed as means +SEM. ???p < 0.001. Student?s t test. (C) Maximum intensity projection of the tumor vasculature in the xenografts with or without PF-502 (1 ?M for 72 h) treatment at 3 dpi, showing the spatial distribution of endothelial cells in neo-vessels. (n = 6). Scale bars: 20 ?m. (D) The density of endothelial nuclei in neo-vessels of the xenografts with or without PF-502 (1 ?M for 72 h) treatment at 3 dpi. (E) GSEA showing the negative enrichment of the HALLMARK_PI3K_AKT_MTOR_SIGNALLNG (NES = ?2.10; FDR q = 0.0), HALLMARK_MTORC1_SIGNALING (NES = ?2.55; FDR q = 0.0), HALLMARK_G2M_CHECKPOINT (NES = ?2.31; FDR q = 0.0) and HALLMARK_E2F_TARGETS (NES = ?2.34; FDR q = 0.0) gene sets in pHUVECs after treatment with PF-502 (0.15 ?M for 12 h). (F) KEGG analysis of the downregulated genes (1364) in pHUVECs after being treated with PF-502 (0.15 ?M for 12 h) (p < 0.005). (G) Cell cycle analysis of the pHUVECs with or without PF-502 (0.15 ?M for 12 h) treatment is based on the G2 M and S gene sets.