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. 2018 Mar 21;9:567. doi: 10.3389/fimmu.2018.00567

Figure 2.

Figure 2

Dectin signaling at fungal synapse (FS). (A) Representative confocal micrographs of macrophage-like THP-1 cells, unstimulated (0 min) or stimulated with C. albicans hyphae for 10, 20, or 30 min to observe the recruitment of Dectin-2 (green), Syk (blue), and autoimmune regulator (AIRE) (red) to the FS. (B–G) Each dot represents a cell used for the measurement indicated. The horizontal bar in each column denotes the mean. *p < 0.05 as determined by the t-test. (B,C) Amounts of AIRE (B) and Syk (C) on the cell surface. (D,E) Amounts of AIRE (D) and Syk (E) at the FS (conjugated–unconjugated). (F,G) Synaptic co-localization of AIRE with Syk (F) or of Syk with AIRE (G). (H) Western blotting analysis of AIRE in cytoplasmic and nuclear fractions from macrophage-like THP-1 cells, unstimulated (unst) or stimulated with hyphae for 10, 20, or 30 min. (L) Ladder, GAPDH and Lamin B1 was used as loading control. (I) Lysates from macrophage-like THP-1 cells, unst or stimulated with hyphae for 30 min, were immunoprecipitated with AIRE (left panel) or Dectin-2 antibodies (right panel) and with a control antibody (IgG). The immunoprecipitated products were probed for AIRE, Dectin-2, and Syk as indicated. Representative confocal micrographs of 30 cells counted in 3 independent experiments. Scale bar = 5 µm.