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. Author manuscript; available in PMC: 2014 Jan 15.
Published in final edited form as: Cancer Res. 2012 Dec 5;73(2):897–907. doi: 10.1158/0008-5472.CAN-12-2130

Figure 2. Activated hepatic myofibroblasts exhibit alterations in their Bcl-2 protein profile and are sensitized to navitoclax by Bax.

Figure 2

Whole cell lysates were prepared from quiescent hFB, hepatic myofibroblasts LX-2 and hCAF as well as quiescent rat fibroblasts (rFB), rat cancer associated fibroblasts (rCAF) and the malignant erbB-2/neu transformed rat cholangiocyte cell line (BDEneu) that is employed in the described in-vivo model of cholangiocarcinoma. Cell lysates were subject to immunoblot analysis of Bcl-2 proteins. Except where indicated by white lines, all lanes were adjacent on the membranes; in some cases, additional lanes originally run between those shown are omitted for clarity (panels A and B). All full-length blots/gels are presented in Supplementary Figure 6. hFB, LX-2 and hCAF cells were grown to approximately 50% confluency on glass chamber slides and treated with navitoclax (1μM) for the indicated time. Cells were then analyzed by fluorescence microscopy using a conformation-specific antibody (6A7) against activated Bax. Bax positive cells are plotted as percentage of all cells (panel C; mean ± SEM; n=4; ** p≤0.01). Wild-type LX-2 cells as well as stably transfected shBax and shBak LX-2 cells were treated with navitoclax (1μM, 24h) and apoptosis was assessed by DAPI staining and fluorescence microscopy (panel D upper graph; mean ± SEM; n=4; ** p≤0.01) as well as fluorometric measurement of caspase 3/7 activity (panel D lower graph; mean ± SEM; n=6; ** p≤0.01)