Skip to main content
. 2012 Dec 20;7(12):e52258. doi: 10.1371/journal.pone.0052258

Figure 3. Chn1 and Chn2 knockdown affect Rac1 activity and cyst morphology.

Figure 3

MDCK cyst expressing shLuc, shChn1, and shChn2 together with Raichu-Rac1 were stimulated with HGF for 30 minutes. And then Rac1 activity of the cells in the cyst wall was calculated at the apical membrane (A) and the lateral membrane (B). Red and green lines indicate the average of FRET/TFP ratios and SD, respectively. *P<0.05. (C) MDCK cysts expressing shLuc, shChn1, and shChn2 were treated with HGF for 2 days and imaged. Shown are the averages of three independent experiments with SD. The numbers of scored cysts are: Experiment (Exp.) 1 [n = 75 (shLuc), 62 (shChn1), 72 (shChn2)]; Exp2 [n = 80 (shLuc), 55 (shChn1), 66 (shChn2)]; Exp3 [60 (shLuc), 70 (shChn1), 61 (shChn2)]. *P<0.05. (D) Representative images of MDCK cysts expressing shLuc (upper), shChn1 (middle), and shChn2 (bottom) together with Raichu-Rac1. Bright-field images (left column), TFP images (middle column) were obtained with an inverted confocal microscope, and FRET/TFP ratio images (right column) were calculated. Bars indicate 10 µm. White arrowheads indicate low Rac1 activity at the apical domain. Red arrowheads indicate luminal cell accumulation showing high Rac1 activity through the whole plasma membrane.