Fig. 4: Impact of PFKFB3 on CRPC cell migration and metastasis.

A Schematic representation of the cellular morphological and behavioral changes following shPFKFB3 in DU145 cells, highlighting reduced migratory and invasive capacities. B Transwell migration assay showing reduced migration in DU145 cells with shPFKFB3 and increased migration in C4-2 cells with OE-PFKFB3. C Transwell invasion assay demonstrating inhibited invasion in DU145 cells with shPFKFB3 and enhanced invasion in C4-2 cells with OE-PFKFB3. D Quantification of migrated cells from (B), based on Transwell assay results. E Quantification of invaded cells from (C), based on Transwell assay results. F Wound healing assay showing reduced migration in DU145 cells with shPFKFB3 and enhanced migration in C4-2 cells with OE-PFKFB3. G Quantification of cell migration in the wound healing assay from (F). H Electron microscopy analysis showing structural changes in DU145 cells after shPFKFB3, indicating alterations in cellular morphology. I Western blot analysis of Cortactin and N-WASP expression in DU145 cells with shPFKFB3, revealing decreased levels of these key cytoskeletal regulators. J Densitometric quantification of Cortactin and N-WASP levels from (I), normalized to GAPDH. K Schematic representation of organ-specific metastasis in nude mice following orthotopic injection of RM-1 cells with OE-PFKFB3. L Gross images of the heart, liver, spleen, lung, kidney, prostate tissues, and metastatic lesions dissected from mice three weeks after establishing an orthotopic PCa model using RM-1 cells overexpressing PFKFB3. The metastatic lesions were observed in the abdominal cavity of the mice. M H&E staining of the prostate, liver, and metastatic lesions in nude mice after orthotopic injection of RM-1 cells overexpressing PFKFB3. The data were presented as the mean ± SD values. ^*P < 0.05; ^**P < 0.01, ^***P < 0.001 by One-way ANOVA. Related to Fig. S5.