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.