Fig. 5

RHPS4 induces reduction of CHK1 and RAD51 and determine S-phase blockage in GSCs. Analysis of proteins involved in DNA damage response and checkpoint activation in patient derived-GSCs (a). Representative blots for CHK1, RAD51 and PCNA in patient derived-GSCs and U251MG (−Adh and -Sph) (b). CHK1 and RAD51 protein level and gene expression in all the cell lines analyzed (c, d, e, f). Data represent mean ± s.d. (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 (Student’s t-test). Assessment of the BrdU-incorporating cells in samples exposed for 4 days to RHPS4. Cells were pulsed for 3 h with BrdU and, after RHPS4 release, cells were chased for additional 24 h (cell were fixed after 4, 6, 8 and 24 h) (g). Quantification of BrdU positive (BrdU+) and negative (BrdU-) cells in untreated (h) and RHPS4-reated cells (i). Note the total depletion of BrdU+ observed over-time in RHPS4 treated samples