Figure 8.

Maneuvers that reduce cysts inhibiting HSP70. (A, B) Western blot of WT and del4 cholangiocytes treated with VER155008. WT and del4 cholangiocytes were grown on Matrigel and monitored for 1 week. (C, D) Cystic cells were treated with HSP70 inhibitor (VER155008). A significant difference from WT in cyst area was observed in HSP70 inhibitor treated cholangiocytes. Localization of CFTR to (E, F) apical vs (G, H) basolateral membranes. CFTR (red) and WGA (green) in WT or del4 cholangiocytes treated with VER155008 (10 μM). Pearson's correlation coefficient R. (F) CFTR (red) and (H) Na⁺/K⁺ ATPase (green) in WT or del4 cholangiocytes. Cells were grown on Transwell membranes. Z-stacks of confocal images acquired at 0.4 μm using a 63× oil objective but representative images of the apical location for WGA vs. the more basolateral image for Na⁺/K⁺ ATPase. A portion of the image demarcated by a white box was enlarged on the right. Scale bar = 10 μm. All data are expressed as mean ± SEM. P values were determined by Student’s t test. For quantification of colocalization, 2–3 random areas were measured along a single z-plane as depicted by the arrows, from 3 individual slides for each group. Number of Western blot experiments represented in the figure (dots) are from independent 10-cm dishes of cultured cells. Number of cysts represented in the figure (dots) are from individual cysts. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001.