Figure 4

Effects of co-treatment on growth of multicellular spheroids (MCS). (A) Multicellular spheroids were prepared as described in Materials and methods. After treatment with indicated concentrations of panobinostat, cisplatin or with combination of both for 24 hours, medium was replaced and spheroids were cultivated under standard normoxic conditions. MCS size was measured every second day for 10 days and on day 10 photographs were made. (B) MCS were incubated with indicated panobinostat concentrations. MCS size was measured every second day over 10 days and the relative cross-sectional area (single time-point values normalized to MCS size on day of treatment = day 2) were determined. Group comparisons were performed with Two-way ANOVA and Bonferroni post-hoc analysis. The significances for panobinostat (32 nM) vs. DMSO are shown. (C) After treatment with single drugs or with combination of both the MCS size was determined as already described. The significances for cisplatin vs. cisplatin + panobinostat are shown. Pano, panobinostat; Cis, cisplatin; ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001.