Figure 3
From: IL-12 and IL-27 regulate the phagolysosomal pathway in mycobacteria-infected human macrophages
Treatment of BCG-infected macrophages with IL-12 and sIL-27R increases expression of CD63. Macrophages were treated with IL-12, sIL-27R, or their combination for 6 h and then infected with SYTO-9® -stained BCG for 48 h. (A) The cultures were subsequently fixed with 4% PFA, permeabilized, and labeled with anti-CD63 antibody (red). Representative images from three experiments are shown (scale bar = 10 μm). Arrows indicate the association of CD63 with BCG. (B) Percent bacterial association was calculated as described in the Methods section. These data are representative results from three independent experiments. (C) Cells were lysed to collect whole-cell lysates for immunoblot analysis (upper panel). CD63 or actin was labeled as described in the Methods Section. An image representative of two experiments is shown. The ratio of CD63/actin band intensity was expressed relative to medium alone (lower panel). Data plotted here are the result of two combined experiments. (B, C) A student’s t test was used to establish statistical significance in the 95% confidence interval between individual sample groups as indicated.