Inhibition of p-ERK with UO126 decreases Snail and reverts EMT independent of proteasomal degradation.

<p>(A) Expression of p-ERK, ERK and Snail was analyzed by Western blot analysis in MCF-7 Neo and MCF-7 Snail cells treated with DMSO control (Ctrl) or UO126 for 30 min, 2 h, 6 h, and 24 h. (B) MCF-7 Neo and MC-7 Snail cells <u>were treated with either DMSO (control) or UO126 and stained with DAPI</u>. Cell morphology and integrity were analyzed by merging DAPI immunofluorescence imaging with brightfield microscopy. (C) Expression of E-cadherin and vimentin in cells treated with DMSO Ctrl or UO126 was analyzed by Western blot analysis. (D) MCF-7 Snail were treated for 6 h and 24 h with DMSO Ctrl, UO126 and UO126+MG132. Cell lysates were analyzed by Western blot analysis. (E) Migration and (F) adhesion assays was performed on MCF-7 Neo and MCF-7 Snail <u>cells</u> treated with DMSO Ctrl or UO126. β-actin was utilized as a loading control for Western blot analysis; DAPI was used to identify the nuclei in immunofluorescence analyses. <u>Maginification 40X</u>. <u>Statistical Analysis was done using ANOVA and Tukey's Multiple Comparison as Post Hoc (**p≤0.01, ***p≤0.001). Values were expressed as mean ± S.E.M (N = 3)</u>. Results are representative of at least three independent experiments.</p>