Research Article

The Autoantigenic Proinsulin B-Chain Peptide B11-23 Synergises with the 70 kDa Heat Shock Protein DnaK in Macrophage Stimulation

Figure 3

Competition of complex formation between DnaK and RCMLA by insulin B-chain peptides. DnaK was incubated with RCMLA in the absence or presence of increasing concentrations of the peptides B11-23 or B18-30. DnaK-RCMLA complexes were separated from free DnaK and DnaK-peptide complexes by native PAGE and visualised after blotting by the use of anti-DnaK antibodies and ECL. (a) Result of a representative blot. (b) The strengths of the resulting signals were quantified by a luminescence detection system and are shown as relative luminescence intensity setting the signals of DnaK-RCMLA samples in the absence of competing peptide as 100%. Data show means + SD from 3 separate experiments. and compared to the luminescence intensity of the DnaK-RCMLA sample in the absence of peptide. Peptides at 2.5 μM correspond to 3.7 μg/ml of peptides, 10 μM correspond to 14.6 μg/ml of peptides, and 40 μM correspond to 58.6 μg/ml of peptides.
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