Supplementary Figure 1 from Constitutively Active ErbB2 Regulates Cisplatin-Induced Cell Death in Breast Cancer Cells via Pro- and Antiapoptotic Mechanisms

Abstract

<p>Supplementary figure 1: Effects of cisplatin treatment on expression of ΔNErbB2 and NHE1, and subcellular localization of cytochrome c. MCF-7 vector (V) and ΔNErbB2 (ΔN) cells were washed free of tetracycline 48 h before the experiment, and exposed to cisplatin (25 μM) or control conditions for 18 h. Cells were subsequently processed for immunoblotting (A), immunofluorescence analysis (B) or cell surface biotinylation (C-D). A. ΔNErbB2, top panel shows representative immunoblots, lower panel shows ΔNErbB2 protein level relative to that in vector cells under control conditions. B. Subcellular localization of cytochrome c. Nuclei are stained with DAPI. C. Representative immunoblots of NHE1 in total lysate fractions (TLF), intracellular fractions (ITF) and biotinylated fractions (BF) in MCF-7 vector (V) and ΔNErbB2 (ΔN) cells. F. Top panel shows representative immunoblots of NHE1 in TLF and BF in native MCF-7 cells, lower panel shows NHE1 protein level relative to that in native MCF-7 cells under control conditions. Data are representative of 3-5 independent experiments for each condition. Western blot data are shown as means with SEM error bars. * indicates comparison of the value in cisplatin treated to that in the respective cell line control, # indicate comparison of values in vector- and ΔNErbB2 cell lines under same treatment. */#, **/##, ***/### and ****/#### indicate p < 0.05, p < 0.01, p < 0.001 and p < 0.0001, respectively (C, one-way ANOVA with Bonferroni post-test) (A, two-way ANOVA with Bonferroni post-test).</p>