Combinatorial effect of Ageritin and TMZ on ZAR and NULU glioblastoma cell lines. (A,B) Western blot analysis of MGMT protein of long-term treated ZAR and NULU cells with Ageritin 0.5 µM for 24, 48 and 72 h. Normalisation was performed with housekeeping gene Actin. Densitometric analysis of protein levels represent the means ± SEM of 3 individual determinations. Data are expressed as fold change over control-treated cells. (C,D) Pretreatment of NULU and ZAR cell lines with Ageritin 0.5 µM followed by TMZ 10 µM for 24 h. Cells treated with TMZ 10 µM for 24 h and Ageritin 0.5 µM for 24 and 48 h as single agents, and untreated cells were used as the control. Data are reported as mean ± SEM of 3 individual determinations. Unpaired t-test, p-value < 0.05. According to GraphPad Prism 7, * or # p-value 0.01 to 0.05 Significant, ** or ## p-value 0.001 to 0.01 (very significant), *** p-value 0.0001 to 0.001 extremely significant, **** or #### p-value < 0.0001 extremely significant.