Fig. 3: Impact of PFKFB3 on cell cycle progression, apoptosis, and organelle integrity in CRPC.

A Schematic representation of cell cycle changes following shPFKFB3 in DU145 cells and OE-PFKFB3 in C4-2 cells. B Flow cytometry analysis showing the impact of shPFKFB3 in DU145 cells and OE-PFKFB3 in C4-2 cells on apoptosis. C Quantification of the cell cycle data. D Quantification of apoptosis, expressed as the percentage of apoptotic cells, based on flow cytometry data. E TUNEL assay results illustrate DNA fragmentation in DU145 cells with shPFKFB3 and C4-2 cells with OE-PFKFB3. F Quantification of DNA fragmentation in DU145 and C4-2 cells, based on TUNEL assay. G Western blot analysis of cell cycle and apoptosis markers in DU145 and C4-2 cells following shPFKFB3 and OE-PFKFB3. H Densitometric quantification of cell cycle and apoptosis markers from (G), normalized to GAPDH. I Electron microscopy showing organelle damage in DU145 cells with shPFKFB3. pseudopodia (PS), nucleus (N), nucleolus (Nu), mitochondria (M), dilated rough endoplasmic reticulum (RER) with ribosomes attached to the membrane, golgi apparatus (GO), lipid droplets (LD), autophagolysosomes (ASS). J Laser confocal microscopy showing mitochondrial damage in DU145 cells following shPFKFB3. K Laser confocal microscopy showing lysosomal fragmentation in DU145 cells with shPFKFB3. L Laser confocal microscopy showing endoplasmic reticulum dilation in DU145 cells after shPFKFB3. M Laser confocal microscopy showing Golgi apparatus fragmentation in DU145 cells with shPFKFB3. N Schematic illustration summarizing organelle damage induced by shPFKFB3 in DU145 cells. O ROS analysis in DU145 and C4-2 cells showing the effects of shPFKFB3 and OE-PFKFB3 on cellular oxidative stress. P Seahorse OCR analysis in DU145 and C4-2 cells showing the effects of shPFKFB3 and OE-PFKFB3 on mitochondrial respiration. Q Quantification of OCR data from (P), normalized to 5000 cells. R Seahorse ECAR analysis in DU145 and C4-2 cells illustrating the effects of shPFKFB3 and OE-PFKFB3 on glycolytic activity. S Quantification of ECAR data from (R), normalized to 5000 cells. The data were presented as the mean ± SD values. ^*P < 0.05; ^**P < 0.01, ^***P < 0.001, ^****P < 0.0001, ns no significant difference by One-way ANOVA. Related to Figs. S3 and S4.