Fig. 2
C. albicans induces pro-inflammatory cytokines in whole-brain and BV-2 cells. C57BL/c mice were challenged i.v. with 25,000 CFU of C. albicans after which whole brains were harvested at the indicated days for analysis by real-time quantitative PCR (RT-qPCR), western blot, or ELISA. a, b Nuclear factor kappa B (NF-κB) expression as assayed by RT-qPCR (a) or western blot (b; p65 subunit) over 14 days. c Densitometric analysis of the data from b. d–f IL-1β, IL-6, and TNF cytokine levels from whole-brain homogenates as assessed by ELISA. g BV-2 cells were seeded for 6 h in 24-well plates (1 × 105 cells per well) and then incubated with C. albicans (200 viable cells per well) for 16 h after which secreted IL-1β, IL-6, and TNF were quantitated by ELISA. h BV-2 cells were seeded same as above and incubated with lysates of C. albicans (equivalent 200 viable cells per well), irradiated C. albicans (200 cells per well), secreted aspartic proteases (SAP, 1 μM), or inhibited secreted aspartic proteases (1 μM) for 16 h and the same cytokines were quantitated by ELISA (n = 4. mean ± S.E.M, ns: not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, using two-tailed Student’s t-test (g) or one-way ANOVA (a, c–f, h) followed by Dunnett’s test for multiple comparison). Data are shown as representative of two independent experiments
