FIG 7.

Autophagy activation by trehalose drives CD63-independent LMP1 release. (A) Confocal microscopy of GFP-LMP1-transfected control or CD63 CRISPR cells following trehalose treatment. Vacuoles were stained with MDC before live-cell imaging. Size bars, 20 μm. N, nuclear compartments. (B) Immunoblot analysis of cytoplasmic lysates from HEK293 control or CD63 CRISPR cells following transfection of GFP or GFP-LMP1 and treatment with trehalose for 24 h, with equal protein mass loaded. (C) Immunoblots of corresponding cell-derived EVs, with equal volumes loaded. Blots are representative images from repeated independent experiments. Quantitation of LMP1 secreted following trehalose treatment was normalized to parental cell secretion over multiple experiments.