a, Time course of flow-cytometry analysis of ADAM10 following lysosomal inhibition with ammonium chloride (NH4Cl, 20 mM), chloroquine (CQ, 50 μM) or PBS as a control; n = 3. b–d, Western blot analysis of ADAM10 and SQSTM1 following lysosomal inhibition with NH4Cl, CQ or bafilomycin (BAF, 10 nM). Shown are a representative western blot from four independent experiments (b), quantification of ADAM10 levels (n = 5) (c) and quantification of SQSTM1 levels (n = 3) (d) at 24 h after inhibition. e, f, Representative histogram (e) and quantification (f) of cell-surface EpCAM in BAF-treated A549 cells; n = 3. g, Time course of flow-cytometry analysis of EpCAM following treatment with NH4Cl or CQ; n = 4. h, i, ADAM10, P4D1 and actin levels following proteasomal inhibition with the chemical compound MG132. Shown are a flow-cytometry time course of cell-surface ADAM10 levels following MG132 treatment (h) and a representative western blot from three independent experiments (i); n = 3. Measurements were taken from distinct samples and graphs show mean ± s.e.m. a, c, d, f–h, One-way ANOVA with Dunnet’s post-test compared with PBS treatment or time 0.