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. 2013 Jun 28;8(6):e67263. doi: 10.1371/journal.pone.0067263

Figure 2. Upregulation of pro-IL-1ß and conversion to mature IL-1ß following exposure to A2E.

Figure 2

(A) Western blot showing upregulation of pro-IL-1ß following stimulation with A2E. ARPE-19 cells were prestimulated with IL-1α and then treated with 0 and 10 µM A2E for a period of 24 hours. Cells were lysed (on ice in the presence of a protease and phosphatase inhibitor) and equal amounts of lysate (assessed via BCA protein assay) underwent western blotting. Membranes were probed with anti-human IL-1ß, which detects both the pro and mature form of the cytokine. Staining with ß-tubulin was used to confirm that comparable amounts of lysate were used. (B) Western blot of cell lysate showing both upregulation of pro-IL-1ß, with some conversion to mature IL-1ß, following stimulation with A2E. (C) Western blot of cell culture supernatant showing increasing amounts of mature IL-1ß in the supernatant, with increasing concentration of A2E.