Figure 3
Tissue lysate array analyses of Hsp27, pSer15, pSer78 and pSer82. Equal amounts of total proteins from normal tissues (n = 10), HER-2/neu positive (n = 10) and HER-2/neu negative (n = 10) breast tumor tissues were manually arrayed on the PVDF membrane in duplicates. Arrays were probed with the respective antibodies and the chemiluminescent signals generated were detected as described in Figure 2. The relative phosphorylation levels (pSer/Hsp27) of Hsp27 at Ser82 (A), Ser15 (B), and Ser78 (C) of each tissue subtype were expressed as the ratio of spot intensity probed with phosphorylation site-specific antibody to the spot intensity probed with anti-Hsp27. The overall average level of pSer/Hsp27 with the standard deviation (± SD), as presented (histograms with error bars) was calculated from 10 cases of each tissue subtype (normal, HER-2/neu negative and HER-2/neu positive tumor tissues). * Significant differences of three groups (normal, HER-2/neu positive and -negative tumors) were calculated using ANOVA. *p < 0.05.