Fig. 3.

Treatment with ABT263 lacks senolytic effect in AR ligand-induced cellular senescent PCa LNCaP cells. LNCaP cells were first treated for 72 h with 1 nM R1881, 10 μM ENZ, or 0.1% DMSO as solvent control. After that, the AR ligands were removed. Fresh medium with 0.1% DMSO or 1 μM ABT263 was added and further incubated for additional 72 h. a Growth of LNCaP cells was analysed by crystal violet staining and OD 590 nm measurement. Values obtained from day 0 were set arbitrarily as 1. Line graphs are shown as mean ± standard deviation (n = 2). Red circles indicate the time point where protein extraction was performed. b The protein extraction was performed after 24 h treatment with ABT263. Detection of full-length PARP (PARP FL), cleaved PARP (c-PARP), RIP3, and phosphorylated RIP3 (p-RIP3) was performed by Western blotting and normalized to β-Actin levels. Upper and middle numbers indicate normalized p-RIP3 and RIP3 band intensities. Lower numbers indicate the ratios of p-RIP3 versus RIP3 levels. c Quantification of fold c-PARP levels normalized to β-Actin from Western blotting data. Values obtained from DMSO + ABT were set arbitrarily as 1. Bar graphs are shown as mean ± SEM (n = 3). d Percentage of SA-β-Gal positive stained cells after 24 h treatment with ABT263. Bar graphs are shown as mean ± SEM (n = 3)