Figure 2
Western blotting of Hsp27, pSer15, pSer78 and pSer82. Twenty μg proteins from each of 4 HER-2/neu positive and 4 -negative tumour samples were separated by 10% SDS-PAGE and transferred onto PVDF membranes. After blocking, the membranes were incubated with the respective primary antibodies (anti-Hsp27, anti-pSer15, anti-pSer78 and anti-pSer82), followed by hybridization to HRP-conjugated secondary antibody. The chemiluminescent signals emitted were captured with the MULTI-GENIUS Bio-Imaging System and signal intensities were analyzed using the GeneTools software (Syngene). The relative phosphorylation levels of pSer15, pSer78 and pSer82 presented (histograms, right) are the respective ratios of signal intensity probed with phosphorylation site-specific antibody to signal intensity probed with anti-Hsp27, for each of the three pSer residues. Data with ± SD (standard deviation) are expressed as the average of triplicate experiments. *p < 0.05 (Student t-test).