Figure 1.

Construction of stably transformed cancer cell lines subjected to Hsp70 gene silencing. (A) HeLa, A549 and MCF-7 cells were transfected with pcDNA3-siRNA-Hsp70 plasmid. Stably transfected cells were isolated using G418 as a selection marker. (B) Cell lines without Hsp70, exhibited a different phenotype similar to that of mesenchymal cells. (C) MCF-7, MCF7-siRNA-Hsp70, A549, A549-siRNA-Hsp70, MCF7 and MCF7-siRNA-Hsp70 cells were heat treated (+) at 41.5°C for 120 min with 90 min recovery at 37°C. RIPA cell extracts were subjected to SDS-PAGE and western blot analysis using monoclonal antibody specific for the inducible Hsp70 and α-tubulin for normalization. (D) Identification of Hsp70 by flow cytometry.