Figure 6. Endogenous Crumbs3a and exogenous Snail expression correlate inversely.

A) Epifluorescent images showing loss of tight (Crb3a) and adherens (E-cadherin) junctions after induction of Snail. Inducible Snail-V5 MDCK cells were grown on transwell filters in the absence (−Dox) or presence (+Dox) of 100 ng/ml doxycycline for 4 days, fixed and immunostained with Crb3a, V5 and E-cadherin antibodies. White bars, 10 μm. B) The same cells were lysed after growth in absence (−) or presence (+) of 100 ng/ml doxycycline for the indicated number of days, or C) for the indicated number of days following doxycycline washout (following initial induction for 7 days). Cell lysates were immunoblotted with indicated antibodies. Graphs (right of each figure) show quantification of immunoblot band intensities as determined by densitometry (see Methods), normalized to β-actin, and reported relative to the maximum protein expression level particular to that experiment.