Figure 6. Influence of HSP90 inhibitor on p53 expression at the transcription and translation levels.

(A) Expression of p53 and GAPDH mRNA as a control was examined with semi-quantitative RT-PCR. JMN-1B cells infected with Ad-p53 (3 × 10⁴ vp/cell) and/or treated with 17-AAG (1 μM) or 17-DMAG (0.1 μM) were subjected to mRNA extraction at the time indicated and the RT-PCR was conducted with different PCR cycles as indicated. GAPDH expression was firstly adjusted and then p53 expression was tested. Relative intensity of PCR bands subtracted with background intensity was estimated with ImageJ software and expressed with arbitrary units. (B) Expression of p53 and actin as a control tested with Western blot analysis. JMN-1B cells treated as above were subjected to cell lysis at the time indicated. (C) JMN-1B cells were infected with Ad-p53 (3 × 10⁴ vp/cell) and/or treated with 17-AAG (1 μM) for the indicated time. Expression of p53 and actin as a control was analyzed with Western blot analysis.