Supplementary Figure 1: Proteasome inhibitor MG132 prevents appropriate T cell activation. | Nature Immunology

Supplementary Figure 1: Proteasome inhibitor MG132 prevents appropriate T cell activation.

From: Integrative proteomics reveals an increase in non-degradative ubiquitylation in activated CD4+ T cells

Supplementary Figure 1

a, Flow cytometry measurement of surface CD44 mean fluorescence intensity (MFI) fold change during TCR stimulation in CD4+ T cells rested or restimulated for 4 h with CD3+CD28 antibody coated beads (1:1 cell:bead ratio). Restimulated cells were untreated or treated with MG132 or cycloheximide (CHX) for the entire 4 h stimulation. b, Flow cytometry measurement of surface CD3ε-γ MFI fold change during TCR stimulation in CD4 + T cells, as described in (a). a, b MFI fold changes are normalized to intensity in unstimulated T cells within the same experiment. Data are compiled from 2 independent experiments comprising 8 mice, mean shown ± SEM. P values calculated by one-way ANOVA with Holm–Sidak test for multiple comparisons, **P < 0.01***P < 0.001, ****P < 0.0001. c, Flow diagram showing expression of surface CD69 in CD4+ T cells rested or restimulated for 4 h using CD3+CD28 antibody coated beads (3:1 cell:bead ratio). A representative plot of activation achieved in three WCP experiments is shown (minimum 65% activation across three experiments). Previously gated on live singlets, CD3 + CD4 + . d, Gating strategy for flow cytometry analysis.

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