Generation of glyceraldehyde-derived advanced glycation end-products in pancreatic cancer cells and the potential of tumor promotion

Figure 3 Western blotting analysis of HSP70 and HSP27. PANC-1 cell lysates (15 μg of proteins/lane) were loaded on a 40-150 g/L polyacrylamide gradient gel. A: Proteins on the polyvinylidene difluoride (PVDF) membrane were probed with anti-HSP70 and anti-GA-3 phosphate dehydrogenase (anti-GAPDH) antibodies. We found four high-molecular-weight (HMW) complexes of HSP70s only in PANC-1 cells treated with GA. Their MWs were 138, 155, 184 and 244 kDa; B: Expression levels of the monomer HSP70 were normalized with GAPDH; C-F: The 138 kDa HSP70/total HSP70, 155 kDa HSP70/total HSP70, 184 kDa HSP70/total HSP70, and 244 kDa HSP70/total HSP70 ratios; G: Proteins on the PVDF membrane were probed with anti-HSP27 and anti-GAPDH antibodies. A HMW complex with a MW of 54 kDa appeared only in PANC-1 cells treated with GA; H: Expression levels of the monomer HSP27 were normalized with GAPDH; I: The 54 kDa HSP27/total HSP27 ratio; A and G: WB was performed for three independent experiments. GAPDH was used as a loading control. B-F, H, I: Data are shown as mean ± SD (n = 3). P values were based on Dunnett’s test. ^aP < 0.05, ^bP < 0.01 vs control. GA: Glyceraldehyde.