A. Gene set enrichment analysis of RNA-seq data from shCont and shCK5-22 MDA-MB-468 (left) and BT20 (right) cells identified Hallmark EMT as a significantly enriched pathway in both cell lines. B. Heatmaps of select EMT genes were generated from triplicate RNA-seq samples of shCont and shCK5-22 in MDA-MB-468 (left) and BT20 (right) cells, normalized to each gene.
C. Cell lysates were collected from shCont and shCK5 (left) or shCK17 (right) BT20 and MDA-MB-468 cells (2 shRNA constructs each) and analyzed by immunoblot for CK5, CK17, vimentin (VIM), E-cadherin (E-cad), and ZEB1. β-actin was used as a loading control; E-cad and ZEB1 were run on a separate blot. Protein levels were normalized to β-actin and indicated as fold change over shCont. ND = not detected.