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. Author manuscript; available in PMC: 2013 Apr 3.
Published in final edited form as: Cell Host Microbe. 2012 Mar 15;11(3):277–289. doi: 10.1016/j.chom.2012.01.019

Figure 5. Negative Effect of Rubicon on Host Antifungal Activity.

Figure 5

(A) Rubicon expression suppresses intracellular fungal killing activity. Raw246.7 or BMDMs infected with lenti-GFP or lenti-GFP-Rubicon for Rubicon expression or infected with lentivirus-shRNA-NS or lentivirus-shRNA-Rubicon for Rubicon depeletion were infected with C. albicans for the indicated times and then lysed to determine intracellular C. albicans loads.

(B) Rubicon suppresses intracellular fungal killing activity in a CARD9-binding-dependent manner. Raw246.7 cells expressing vector, Rubicon WT, or its mutants were infected with C. albicans for the indicated times and then lysed to determine intracellular C. albicans loads.

(C–E) Alteration of Rubicon gene expression affects mouse mortality after C. albicans infection. At 48 hr postinjection with Ad-vector (1 × 1013 pfu/kg), Ad-shRubicon (1 × 1012 pfu/kg), or Ad-Rubicon (1 × 1013 pfu/kg) twice i.v. via tail vein, mice were infected with C. albicans (1 × 107 CFU per mouse), and mortality was measured for n = 10 mice per group (C). Serum cytokine levels (D) or C. albicans loads of infected mice (n = 5 per group) in liver, spleen, kidney, and lung (E) were determined at 4 days p.i. with C. albicans (1 × 106 CFU). Additional data including Rubicon expression in mice are shown in Figures S5A and S5B. CFU, colony-forming units.

(F–I) Doxycycline-inducible, macrophage-specific Rubicon expression affects mouse mortality after C. albicans infection. After 7 days of doxycycline treatment, SRA-rtTA/Flag-Rubicon double transgenic mice were infected with C. albicans (1 × 107 CFU per mouse), and mortality was measured for n = 6 mice per group (F). C. albicans loads of infected mice (n = 6 per group) in liver and spleen (G), serum cytokine levels (H), and H&E staining of spleens (I) were determined at 4 days p.i. with C. albicans (1 × 106 CFU). Additional data including Rubicon expression in mice are shown in Figures S5C–S5E. The data are the mean ± SD of values from three experiments. *p < 0.05; **p < 0.01; ***p < 0.001 compared with the control. Scale bars, 200 μm. See also Figure S5.