Figure 3From: Importance of spliceosomal RNP1 motif for intermolecular T-B cell spreading and tolerance restoration in lupusReactivity of rabbit antibodies directed to peptide 131–151 of the U1-70K protein with RNP1-peptides and spliceosomal proteins. (a) The antiserum was diluted 1/20,000 and incubated first for 1 h at 37°C and then overnight at 4°C with increasing amounts of different RNP1-peptides used as competitors in the fluid-phase. The homologous peptide 131–151 was used as control. The mixtures were then added to microtiter plates pre-coated with 2 μM of immunizing peptide 131–151. The results are expressed as the percentage of inhibition of the ELISA reaction measured without competitor peptide. (b) U1-70K, U1-A, U1-C, and SmD1 proteins were subjected to electrophoresis, transferred to nitrocellulose, and colored with Ponceau red (lanes 1–4). The blotted strips were then incubated with the serum (diluted 1/500) from rabbits that received either peptide 131–151 (lanes 5–8) or CFA alone (lanes 9–12). IgG antibodies only were tested. ECL reagents were used to reveal positive reactions.Back to article page