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. 2011 Mar;178(3):983–988. doi: 10.1016/j.ajpath.2010.12.007

Figure 1.

Figure 1

Identification of AIRE association with K17. A: THP-1 cytoplasmic lysate was immunoprecipitated with either rabbit anti-AIRE or rabbit-IgG, resolved on an SDS gel, and then stained with Colloidal Coomassie. A band appearing specific to AIRE below 50 kD was excised, trypsin-digested, and subjected to mass spectrometric analysis. IP, immunoprecipitate. B: Twelve unique tryptic peptides for K17 were identified and their positioning in a schematic of the protein is displayed (left). The arrows demonstrate examples of the K17-specific peptide ion spectra. Assigned β- and γ-ions are labeled. C: RT-PCR analysis of AIRE message in HaCaT cells, with THP-1 monocytes used as a positive control, and COS-1 cells as a negative control, for AIRE expression. Actin is showed as positive control for amplification. D: Confirmatory immunoprecipitation of HaCaT whole-cell lysate with either rabbit anti-human AIRE or rabbit anti-human K17, followed by Western blot (WB) for AIRE (with goat anti-AIRE antibody) or K17 (with rabbit anti-K17 antibody). Isotype immunoprecipitations were performed as negative controls.