Fig. 3: Quantitative fluorescence imaging measuring the effect of 17-AAG on HER2.
From: Chaperone mediated detection of small molecule target binding in cells
a Immunofluorescence images of HER2 protein in MCF-7 neoHER2 cells treated with DMSO or 17-AAG (1 µM) for 4 h. The upper panels show Hoechst 33342 staining of the DNA (blue) indicating the nuclear region, while the lower panels show the HER2 immunofluorescence stain. Scale bar −20 μm. b The change in mean HER2 protein per cell (MCF-7 neoHER2) was calculated for a range of concentrations of 17-AAG and expressed relative to the HER2 level in DMSO treated samples and the no primary antibody control sample. c Chemical structures of HER2 inhibitors: Lapatinib, HY-14674, and TAK-285. d Measurement of the relative HER2 protein levels in MCF-7 neoHER2 cells pre-treated with a range of concentrations of HER2 inhibitors (indicated by color) then treated with a titration of 17-AAG (x-axis) for 4 h. HER2 levels are expressed as normalized fold change in HER2 levels relative to the DMSO treated sample (Norm. Fold Change). e Measurement of the relative HER2 protein levels in MCF-7 neoHER2 cells pre-treated with a titration of each HER2 inhibitor then treated with 2 µM 17-AAG. HER2 protein levels are expressed as the fraction of HER2 present in the co-treatment condition relative to the corresponding concentration of inhibitor only (HIPStA Fold Change). The relative EC50 values were calculated using a four parameter quadratic curve fit in PRISM. Error bars represent standard error of mean (n = 3). Source data are provided as a Source Data file.
