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. 2018 May 31;9:1235. doi: 10.3389/fimmu.2018.01235

Figure 3.

Figure 3

Lack of RhoA halts T-cell immune responses against the encephalitogenic antigen, MOG35–55. (A) Representative histogram of the proliferation of RhoA+/+ and RhoA−/− T-cells in vitro based on their CFSE labeling. (B) Quantification of the proliferation index (fold increase in proliferation in relation to unstimulated RhoA+/+ T-cells) of RhoA+/+ (filled bars) and RhoA−/− (striped bars) T-cells upon MOG35–55 activation for 48 h. Data represents the mean of two independent experiments (RhoA+/+ n = 4, RhoA−/− n = 3). (C) Representative gating strategy for the analysis of cell viability. (D) Quantification of the percentage of necrotic (N), apoptotic (EA + LA) and living (L) cells in RhoA+/+ and RhoA−/− T-cells after 24 h of MOG35–55-activation in vitro. (E–K) Quantification of the amount of IFN-γ, IL-12p40, IL-6, TNF-α, IL-4, IL-5, and IL-17A present in the supernatant of RhoA+/+ (continuous line) and RhoA−/− (dotted line) T-cells after 24, 48, and 72 h in culture in the presence of MOG35–55. Data shown as mean ± SEM *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 [(B) One-way ANOVA, followed by Tukey’s test. (D) Multiple t-test, followed by FDR correction. (E–J) Multiple t-test, followed by FDR correction]. Abbreviation: FSC, forward scatter; SSC, side scatter; PI, propidium iodide; FLICA, fluorescent inhibitor of caspases; L, living; N, necrotic; EA, early apoptotic; LA, late apoptotic; FDR, false discovery rate.