Skip to main content
. Author manuscript; available in PMC: 2013 Nov 16.
Published in final edited form as: Oncogene. 2012 Jul 9;32(20):2543–2554. doi: 10.1038/onc.2012.277

Figure 3.

Figure 3

Effects of chloroquine-mediated lysosomal inhibition on 3D morphogenesis driven by oncogenic PI3K. (A) Monolayer growth curves of PI3K-H1047R cells treated with 10μM (left) or 20μM chloroquine (CQ) (right) in the presence or absence of 50nM Rapamycin (Rap). (B) Left: Phase images of day 7 3D cultures of PI3K-H1047R cells subject to the indicated treatments. Bar, 100μm. Right: Acinar size distribution on day 7 of 3D culture. A total of 200 acini from two independent experiments were measured for each condition. (C) Left: PI3K-H1047R cells were cultured in the presence of absence of 10μM CQ for 7 days. The outgrowths were immunostained with antibodies against Ki67 (green), laminin-5 (red), and DAPI-counterstained (blue). Bar, 50μm. Right: the percentage of Ki67 positive nuclei per acinus was quantified. For each condition, a total of 75 acini from three independent experiments were analyzed. (D) Left: PI3K-H1047R cells were treated with 25nM rapamycin alone or in combination of 10μM CQ. Day 7 outgrowths were immunostained with antibodies against Ki67 (green), laminin-5 (red), and DAPI-counterstained (blue). Bar, 50μm. Right: The percentage of Ki67 positive nuclei per acinus was quantified. For each condition, a total of 75 acini from three independent experiments were analyzed. Results represent the mean ± SEM *** p<0.001, Student's t test.