Fig. 3

PFKFB4-knockdown (KD) impairs cell fitness in ccRCC. A) Cell count detected using CCK-8 in clear-cell renal cell carcinoma (ccRCC) cell lines with PFKFB4-knockdown (KD) by shRNA#1 and shRNA#2 (KD1 and KD2) or scrambled negative control (NC); B) Colony formation in ccRCC cell lines with PFKFB4 silencing or control); C) Proliferation detected using ratio of EDU/DAPI staining in ccRCC cell lines with PFKFB4-KD or NC; Flow cytometry used to detect D) cell cycle profile and E) apoptosis in ccRCC cells with PFKFB4-KD or NC; Transwell assays used to detect F) cell invasion with Matrigel and G) cell migration without Matrigel in ccRCC cells with PFKFB4-KD or NC; Wound healing assay in ccRCC cells with PFKFB4-KD or NC. (All in vitro assays performed in triplicates and at least 3 biological replicates; All comparisons by Student’s test, N = 5; ns = not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001)