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. 2014 Oct 20;211(11):2307–2321. doi: 10.1084/jem.20132349

Figure 6.

Figure 6.

CARD9-mediated H-Ras activation is critical for immune responses triggered by C. albicans yeast and curdlan. (A and B) ELISA results for cytokines TNF, IL-1β, IL-6, and IL-12p40 in supernatants of WT and CARD9-deficient BMDMs, which were stimulated for 6 h with UV-inactivated WT or mnn5 yeast (MOI = 5; A) or hyphae (MOI = 0.1; B). (C and D) ELISA results for cytokines TNF, IL-1β, IL-6, and IL-12p40 in supernatants of WT and CARD9-deficient BMDMs, which were pretreated with or without 5 µM U0126 for 30 min and then stimulated with UV-inactivated mnn5 yeast cells (MOI = 5; C) or plate-coated curdlan (50 µg/ml; D) for 6 h. (E and F) ELISA results for cytokines TNF, IL-1β, IL-6, and IL-12p40 in supernatants of WT BMDMs, which were transfected with siRNA against murine H-Ras and nontargeting control siRNA using Trans IT-TKO transfection reagent (Mirus). Cells were cultured for 48 h after transfection and then stimulated with plate-coated curdlan (50 µg/ml), UV-inactivated mnn5 yeast cells (MOI = 5), plate-coated α-mannans (50 µg/ml), or LPS (100 ng/ml) for 6 h. (G) ELISA results for cytokines TNF, IL-1β, IL-6, and IL-12p40 in supernatants of WT and CARD9-deficient BMDMs, which were pretreated with or without 5 µM U0126 for 30 min and then stimulated with plate-coated α-mannans (50 µg/ml) or LPS (100 ng/ml) for 6 h. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Data are means ± SD of triplicate samples and are representative of three independent experiments.