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. Author manuscript; available in PMC: 2015 May 1.
Published in final edited form as: Gastroenterology. 2014 Feb 11;146(5):1301–1312.e10. doi: 10.1053/j.gastro.2014.02.003

Figure 3. Knockdown of PHLPP expression induces EMT and promotes migration and invasion in CRC cells.

Figure 3

(A) Stable control and PHLPP knockdown SW480 cells were co-stained with E-cadherin and vimentin antibodies and analyzed using fluorescence microscopy. Cell nuclei were stained with DAPI. Scale bars, 10 μm. (B) Cell lysates prepared from stable SW480 knockdown cells were analyzed using immunoblotting. The relative expression levels of E-cadherin and vimentin were quantified by normalizing to tubulin. (C-D) Stable SW480 knockdown cells were subjected to Transwell migration assays (C) using collagen and EGF as chemoattractants and Transwell invasion assays (D) using 5% FBS as the chemoattractant. Each experiment was done in duplicates and three independent experiments were averaged and expressed as mean ± SD (* p<0.01 by two-sample t-tests compared to the control cells). (E) Migration patterns of stable SW480 knockdown cells. The trajectories of 10 randomly chosen cells for each cell line were shown in the graphs. The radius of 5 and 10 μm from the origin is shown as dashed circles for reference. (F) The average distance traveled by stable SW480 knockdown cells during 6 hours of migration. Data represent mean ± SD (n = 10 cells/line, # p<0.05 by two-sample t-tests).