Figure 2.

Forced Snail expression induces a partial EMT in vitro. A: Inducible Snail (imSnail) in mOSE cells show increased SNAIL in the presence of doxycycline (200 ng/mL, 48 h) compared to the control mOSE cells bearing inducible GFP (iGFP), as determined by western blot. CDH1 levels remain unaffected by doxycycline treatment. β-ACTIN was used as a loading control (representative western blot, N = 3). B: imSnail mOSE cells do not show altered expression of EMT markers (Alcam, Krt19) in the presence of doxycycline for 48 h (N = 3, Student t-test). C: imSnail cells have a modestly increased migration in response to doxycycline treatment for up to 48 h (100 ng/mL), compared to iGFP cells, whereas their migration is not significantly different in the absence of doxycycline (control), as determined using a migration assay (N = 3, ANCOVA, R² = 0.93 and 0.99 for untreated iGFP and imSnail cells respectively and 0.98 and 0.95 for Dox treated iGFP and imSnail cells respectively-). ** indicates a significant difference between treatment groups, P < 0.005. D: imSnail mOSE cells do not alter actin cytoskeletal organization in the presence of doxycycline, as determined by ACTIN immunofluorescence (representative image, N = 3). Scale bar = 100 μm. RQ = relative quantity. Experiments were performed using cells under passage number 25 and Snail expression was induced by treating cells with dox for 48 h prior to an experimental readout.