Figure 1
Hsp90 is expressed on the cell surface of breast cancer cells. (a and b) BT-474 (Her2-overexpressing), MCF-7 (ER + , PR + ), and MDA-MB-231 (triple negative) breast cancer cells were stained with 100 µM HS-27 only for 15 minutes (green, control), serially with 100 µM HS-10 for 14 minutes followed by 100 µM HS-27 for 15 minutes (serial), or simultaneously with 100 µM HS-10 and 100 µM HS-27 (simultaneous) for 15 minutes. DNA is stained in blue with Hoechst 33342. Scale bars are 20 µm. Fluorescence was normalized so that the control images had a value of 1. For all experiments, the sample size was n = 3 per group, and p-values were determined using one-way ANOVA followed by Tukey-Kramer post-hoc testing. (c) Western blotting was performed for HSF-1, Hsp70, and Akt expression in BT-474, MCF-7, and MDA-MB-231 cells after treatment with 100 µM HS-27 for 12, 24, or 48 hours. Cropped western blots are shown; full blots can be found in Supplementary Figure S1. Control cells were incubated with DMSO vehicle. Western blots were performed in triplicate.
