Fig. 1

Representative Western blot analysis of cytosolic Hsp70 in CX⁺/CX⁻ (a), 4 T1 ctrl/4 T1 Hsp70 KD (a), H1339 ctrl/H1339 HSF-1 KD, EPLC-272 ctrl/EPLC-272H HSF-1 KD (b) tumor sublines under non-stressed conditions. 20 μg of cytosolic proteins were administered to each lane of a SDS/PAGE and blotted onto nitrocellulose membranes. Hsp70 and ß-actin were detected using primary antibodies, an appropriate HRP-conjugated secondary antibody, followed by visualization with ECL kit. β-actin and recombinant Hsp70 protein (10 ng, 25 ng; data not shown) were used as loading control and for normalization. Normalized Hsp70 values of CX⁺/CX⁻ and 4 T1 ctrl/4 T1 Hsp70 KD (c), H1339 ctrl/H1339 HSF-1 KD and EPLC-272H ctrl/EPLC-272H HSF-1 KD (d, e) cell lysates, as determined by Western blot analysis (c, d) and ELISA of cell lysates (e). Bars represent the mean values of n = 3-4 independent experiments. Significance: *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001