Figure 2

Loss of Hsp90 client proteins. (A) GB cells (U251, A172, and SN19) were treated with or without Antp-TPR peptide at the indicated concentrations, and examined by Western-blot analysis for expression of Hsp90, Hsp70, Hsp27, p53, CDK4, Akt, cRaf, and β-actin using corresponding antibodies. (B) Effect of Antp-TPR peptide on the transcriptional level of Hsp90 client proteins. Total RNA isolated from GB cells treated with or without Antp-TPR peptide at the indicated concentrations were reverse transcribed to cDNA, and then assessed by PCR (inset) or real-time quantitative PCR using specific primers as described in the Materials and Methods section. GAPDH served as an internal control. (C) Effect of Antp-TPR peptide on the Erk1/2 pathway. GB cells were treated with or without Antp-TPR peptide as indicated, and examined by Western-blot analysis for expression of PI3K, Erk1/2, or phosphorylated (p-) Erk1/2 using corresponding antibodies. β-Actin was used as the loading control. All bands of the Western-blot analysis were visualized by chemiluminescence.