Fig. 2

SAHA suppresses cell viability and diminishes the expression stemness-related markers in GSCs. a and b U87MG adherent cells and spheroids were treated with DMSO or different doses of SAHA (0, 3, 6 μM) for various time intervals (1 to 6 days). After treatment, cell viability was assessed using colorimetric MTT assay. c U87MG spheroids were treated with different doses of SAHA as indicated for various time intervals (0, 1, 2, 3 days). After treatment, the cell proliferation was examined by performing cell counting (Trypan blue exclusion) assay. d Photomicrographs of GSCSs treated with different dosages of SAHA were randomly selected in microscopic fields. e and f Cells from dissociated GSCs via trypsinization were grown in soft agar followed by treatment with SAHA at different doses every 48 h. After 14–30 days of incubation at 37 °C, colonies arose from these single cells were photographed randomly and counted in the three fields chosen. The histogram of average colony numbers was obtained by performing the experiment in triplicate; bar, S.D.; *p < 0.05; **p < 0.01. g and h GSCs after 48 h of SAHA treatment were harvested and the protein expression levels of stemness markers were analyzed by Western blotting with indicated antibodies. The protein levels were quantified by imageLab software 5.2.1 (Bio-RAD)