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. 2022 Apr 21;221(6):e202110151. doi: 10.1083/jcb.202110151

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© 2022 Solvik et al.

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Figure 3. — Lysosomal inhibition broadly promotes secretion of autophagy cargo receptors via EVPs. (A) Cell lysate (WCL; left) and 100,000 g EVP fractions (100K; right) from serum-starved HEK293T cells treated with vehicle or 20 nM BafA1 for 16 h were collected and blotted for the indicated proteins (n = 3). (B) Quantification of the proteins in EVP fractions from BafA1 treated cells relative to controls in A. (mean ± SEM; n = 3; *, P < 0.05; **, P < 0.01; ***, P < 0.005). Statistical significance calculated by unpaired, two-tailed t test. (C) WCL (left) and 100K fractions (right) from serum starved HEK293T cells treated with vehicle or 25 µM CQ for 16 h were collected and blotted for the indicated proteins (n = 3). (D) Quantification of the proteins in EVP fractions from CQ treated cells relative to controls in C (mean ± SEM; n = 3; **, P < 0.01; ***, P < 0.005). Statistical significance calculated unpaired, two-tailed t test. (E) Plasma EVPs from mice treated with vehicle or 60 mg/kg hydroxychloroquine (HCQ) for three consecutive days were collected and blotted for the levels of GFP-LC3, p62, and NBR1. (F) Quantification of the indicated proteins in plasma EV fractions from mice treated with vehicle or CQ from E (mean ± SEM; n = 3; *, P < 0.05; **, P < 0.01). Statistics were calculated by unpaired, two-tailed t-test. Source data are available for this figure: SourceData F3.