Figure 2.

(A) The 5′-flanking sequence of the human Hsp70B′ gene (GenBank accession: NT004487). The promoter sequence used in this work is shown in uppercase letters. Nucleotides matching the primers used for PCR amplification are in italics and indicated by dashed underlines. Four consecutive heat-shock-element sequences are underlined. The GATA box sequence is shown by a solid box. The transcription start site is represented by an arrow. (B) GFP expression and (C) cell viability of MCF-7 cells transfected with pHSP-GFP after heat-shock treatment at different temperatures and times. (D) Knockdown of GFP expression by pHSP-shGFP in MCF-7 cells. The indicated RNAi plasmid pHSP-shGFP was co-transfected into MCF-7 cells with the GFP expression plasmid pCMV-GFP at a ratio of 1:1. At 24 h post-transfection, MCF-7 cells were treated with (middle) or without (right) heat-shock at 42°C for 40 min, with untransfected MCF-7 cells used as controls (left).

Abbreviations: GFP, green fluorescent protein; HSP, heat-shock protein; PCR, polymerase chain reaction; sh, short hairpin.