Western Blots
Cells were harvested 48 hours after lipofection using RIPA buffer and total cellular protein was extracted. A BCA assay (Pierce BCA Protein Assay Kit) was used to determine the protein concentration of each sample. 5 ug of lysate from each sample was loaded into a Mini-PROTEAN TGX stain-free precast gel (Bio-Rad) and resolved. Once proteins were resolved, gels were activated on a ChemiDoc (Bio-Rad) imager and were then transferred to a polyvinylidene difluoride (PVDF) membrane. Membranes were probed with mouse Cytokeratin 14 antibody primary antibody (Life Technologies) and HRP conjugated goat anti-mouse secondary antibody (Santa Cruz Biotechnology). Total proteins and chemiluminescence was imaged on the ChemiDoc and Image Lab Software (Bio-Rad) was used to analyze protein abundance.