Fig. 3.

Differential cellular impacts of prostate cancer cell models by SRRM4. (a) The morphology of cells was imaged by Zeiss light microscope, where the scale bars represent 20 μm. Additionally, the DU145(SRRM4), or DuNE, cells and its control cells, DU145(Ctrl), were fixed for IF against F-actin using anti-F-actin antibody conjugated to Phalloidin-iFluor 488 and mounted with DAPI staining mount. Cells were then imaged by confocal microscopy, where the scale bar represents 10 μm. 50 cells were randomly selected to measure the average cell body length in μm in the Image J program and presented on the bar graph. (b) Cells were seeded in a 96-well plate for MTS assays to determine cell viability over a 5-day time course. The DuNE cells and its respective control cells, were seeded in (c) matrigel for 3D BrdU proliferation assays, (d) soft agar for 10-day-long colony formation assays (where colonies >100 μm were counted), or (e) 10cm² dish and treated the next day with Doce (20 nM), CPT (60 nM), vehicle (100% ethanol), or 10 mJ/cm² of UV irradiation for 48 h. The protein lysates were then used to measure c-PARP and vinculin protein levels by immunoblotting assays. Three independent biological replicates were performed for each experiment. All results are presented as the mean ± SD (Student t-test; n = 3; ***denotes p <0.001). IF, immunofluorescence; UV, ultra violet; CPT, camptothecin; Doce, docetaxel; c-PARP, cleaved PARP; OD, optical density.