Figure 7.

Increased luminal cell death in 3D acini upon chemical autophagy inhibition. (A) Indicated cell types were cultured for 6 d on Matrigel upon which 3-MA (10 mM) was added for 2 d. Representative fields of EtBr stained day 8 acini (middle) and corresponding phase contrast images (left) are shown. Bar, 20 μm. Right, day 8 acini were immunostained with α-cleaved caspase-3 (green) and α-laminin-5 (red). DAPI-stained equatorial confocal cross-sections are shown. Bar, 20 μm. (B) After 6 d of 3D culture, 3-MA (10 mM) was added to the indicated cell types; the percentage of acini containing EtBr-positive cells was enumerated on subsequent days. Results are the mean ± SEM of four independent experiments. (C) After 6 d of 3D culture, CQ (20 μM) or BafA (20 nM) was added to the indicated cell types; the percentage of acini containing EtBr-positive cells was enumerated on subsequent days. Results are the mean ± SEM of three independent experiments.