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. 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 4.

Figure 4

Chloroquine-treatment promotes outgrowth of PI3KH1047R cells in 3D culture through activation of ERK/MAPK pathway. (A) PI3K-H1047R cells were cultured in the presence of absence 10μM CQ. FACS analysis of activation specific ERK phosphorylation (pERK) was performed in cells recovered from day 11 3D cultures. (B) Left: Phase images of day 8 3D cultures treated with 10μM U0126, 10μM CQ, or 10μM U0126+10μM CQ as indicated. 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: Day 7 outgrowths were fixed and immunostained with antibodies against pRb780 (green), laminin-5 (red), and DAPI-counterstained (blue). Bar, 50μm. Right: The percentage of pRb780 positive nuclei per acinus was quantified. For each cell type and condition, a total of 100 acini from two independent experiments were analyzed. Results represent the mean ± SEM, * p< 0.05 *** p<0.001, Student's t test. N.S. not significant.