CTLA4 accumulation in USP8-deleted mouse CD4 T cells and secretion via exosomes from USP8-depleted melanoma cells. (A) Representative Airyscan images (two fields of view each) of CTLA4 co-stained with DAPI in USP8 fl/fl (WT, 1–3) and USP8 deleted (∆USP8, 1–3) activated T cells derived from individual mice. Scale bar = 15 µm. (B) A2058 cells were transfected for 72 h with non-targeting (NT1) or USP8, HD-PTP, ALIX, and Syntenin siRNAs. Lysates were collected and the cultured supernatant subjected to TCA precipitation before analysis by SDS-PAGE and western blot. (C) Quantification of CTLA4 secreted into the media for data represented in B. Individual data points from two independent, color-coded experiments are shown. Error bars indicate the range. Source data are available for this figure: SourceData FS3.
Figure S3.

CTLA4 accumulation in USP8-deleted mouse CD4 T cells and secretion via exosomes from USP8-depleted melanoma cells. (A) Representative Airyscan images (two fields of view each) of CTLA4 co-stained with DAPI in USP8 fl/fl (WT, 1–3) and USP8 deleted (∆USP8, 1–3) activated T cells derived from individual mice. Scale bar = 15 µm. (B) A2058 cells were transfected for 72 h with non-targeting (NT1) or USP8, HD-PTP, ALIX, and Syntenin siRNAs. Lysates were collected and the cultured supernatant subjected to TCA precipitation before analysis by SDS-PAGE and western blot. (C) Quantification of CTLA4 secreted into the media for data represented in B. Individual data points from two independent, color-coded experiments are shown. Error bars indicate the range. Source data are available for this figure: SourceData FS3.

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