Fig. 6

Real time qPCR analysis of cytochines IFNγ, IL-12, IL-10, IL-1 β, IL-6 and TNFα (a) or immune checkpoint genes as PD-L1, CTLA-4, TIM-3 and LAG-3 (b) expressed by spheroids untreated (CTR) or treated with selumetinib, atezolizumab or their combinations. Results were normalized to 18S mRNA and analyzed by ΔCt method. One way ANOVA test followed by Tukey’s test were used for statistical analysis. * p < 0.05; **p < 0.01; ***p < 0.001