Macrophage activation in response to <i>mar1Δ</i> requires members of the CLR and TLR families.

<p>BMMs were harvested from the indicated mouse strains (C57BL/6 background, unless otherwise noted) and co-incubated with <i>C</i>. <i>neoformans</i> (Cn) cells at an MOI of 10:1, Cn:BMMs, followed by quantification of TNF-α (pg/ml) in the supernatant by ELISA. (A) Card9 is involved in macrophage activation by <i>mar1Δ</i> cells. Data represent means of 3 replicates from 3 independent experiments (n = 9). **, p < 0.01 WT vs. Card9^-/- BMMs as determined by two-way ANOVA with Sidak’s multiple comparisons test. (B) MyD88 is required for TNF-α production by macrophages in response to <i>mar1Δ</i>. Data represent means of 3 replicates from 3 independent experiments (n = 9). ****, p < 0.0001 WT vs. MyD88^-/- BMMs as determined by two-way ANOVA with Sidak’s multiple comparisons test. (C) Dectin-1 is involved in the response to <i>mar1Δ</i> cells. Data represent means of 3 replicates from 2 experiments (n = 6). ****, p < 0.0001 WT vs. Dectin-1^-/- BMMs as determined by two-way ANOVA with Sidak’s multiple comparisons test. (D) TLR2/4^-/- BMMs do not respond to <i>mar1Δ</i> cells. Data represent means of 3 replicates (n = 3). ****, p < 0.0001 WT vs. TLR2/4^-/- BMMs as determined by two-way ANOVA with Sidak’s multiple comparisons test. (E) TLR4 is not required for the production of TNF-α induced by <i>mar1Δ</i> cells. BMMs were isolated from C3H/HeJ mice with a null mutation in TLR4, and C3H/HeOuJ control mice. Data represent means of 3 replicates from 2 independent experiments (n = 6). **, p < 0.01 C3H/HeOuJ vs. C3H/HeJ BMMs as determined by two-way ANOVA with Sidak’s multiple comparisons test. (F) Macrophage activation by <i>mar1Δ</i> is partially dependent on TLR2. Data represent means of 3 replicates from 3 independent experiments (n = 9). ****, p < 0.0001 WT vs. TLR2^-/- BMMs as determined by two-way ANOVA with Sidak’s multiple comparisons test.