Betamethasone affects splenocytes’ viability and proliferation in vitro. (A) Percentage of viable splenocytes (annexinV PE−, 7aad−) after 24 h of culture with increasing betamethasone concentrations [0.1; 1; 2.5; 5; 10; 50; 100 and 1000 nM] (grey dots), and control condition (white circles). Lines show the mean of n ≥ 4 independent experiments (*p ≤ 0.05 and **p < 0.01, Dunn’s test, Kruskal-Wallis). (B) Sigmoidal non-linear regression of the splenocytes’ viability and the Log2 of the betamethasone concentration to determine the inhibitory concentration 50 (R2 = 0.9887 and IC50 = 2.755 nM). (C) Percentage of viable T cells (annexinV-PE−, 7aad− of CD3+ splenocytes) and B cells (annexinV-PE−, 7aad− of CD19+ splenocytes) after 72 h of culture with betamethasone (bet). White bars correspond to T cells with phorbol 12-miristate 13-acetate and ionomicine (PMA + IO) cultured with complete media (CM), 1 nM, 5 nM or 25 nM bet. Grey bars correspond to B cells with PMA + IO cultured with CM, 1 nM, 5 nM or 25 nM bet. Results presented as range and mean of 5 independent experiments (*p ≤ 0.05, Tukey’s test, Two-Way Anova).