Fig. 9

PFKFB4-knockdown overcomes Sunitinib resistance in ccRCC in vivo. A Xenograft murine models consisting of 8 male BALB/c nude mice per group with subcutaneous implanted Sun-sensitive A498 cells with or without PFKFB4-KD (shRNA#2) under right hind limb with tumor growth monitored over 60-day period and tumor size of < 2500 mm3 as endpoint, with representative tumor image at endpoint (bar = 1 cm) analyzed by two-way ANOVA and B Kaplan-Meier curves of survival of mice, analyzed by Log-rank test; C Representative immunohistochemical staining of factors in tumors from A), with intensity scored semi-quantitatively and statistically compared; D Xenograft murine models consisting of 8 male BALB/c nude mice per group with subcutaneous implanted Sun-resistant 786O cells fed with 20 mg/kg of Sun and/or 120 mg/kg of 5MPN orally by gavage; tumor growth monitored over 60-day period, analyzed by two-way ANOVA and E Kaplan-Meier curves of survival of mice, analyzed by Log-rank test; F Representative immunohistochemical staining CD31 targeting micro-vessels in tumors (red arrows) from D, with relative micro-vessel density (MVD) statistically compared; G Tail vein injection of Sun-resistant A498 cells in 9 mice per group with 5MPN, Sun or combo treatments (Tx); mice monitored for 4 weeks for photon detection, normalized to PreTx CON group; bar figures showing photon change before (PreTx) and after (PostTx) with representative luciferase image showing lung involvement at endpoint of each group H Schematic cartoon of regulatory axis proposed by the current study. (All in vitro assays performed in triplicates and at least 3 biological replicates; ns = not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001)