Fig 1: NF-κB and p38 activation stimulated by eng1Δ/Δ and fgr41Δ/Δ strains. Macrophages and C. albicans strains were co-incubated for 30 min (A–D, I and J) or 4 h (E–H). Macrophage protein was extracted and run on a 12% SDS-PAGE gel, and gels were transferred to PVDF membranes for immunoblotting. All membranes were blotted for tubulin as a loading control, with the primary antibody used at 1:1,000. (A, E) Membranes were blotted for IκBα, and 50 μg of protein was loaded into each well. The primary antibody to IκBα was used at 1:1,000. (B, F) Membranes were blotted for p100/p52, and 20 μg of protein was loaded into each well. The primary antibody was used at 1:1,000. (C, G) Membranes were blotted for p38, and 50 μg of protein was loaded into each well. The primary antibody to p38 was used at 1:1,000. (D, H) Membranes were blotted for phosphorylated p38, and 100 μg of protein was loaded into each well. The primary antibody was used at 1:500. (I) Quantification of D, including the positive control (stimulatory laminarin). Levels of phospho-p38 are significantly increased in the positive control condition. ***P < 0.001 by one-way ANOVA, ns = not significant. (J) Quantification of D, excluding the positive control condition. Both eng1Δ/Δ- and fgr41Δ/Δ-challenged macrophages show a significant increase in the production of phospho-p38 over untreated macrophages (MΦ only). Challenge with the fgr41Δ/Δ mutant also causes significantly more phospho-p38 production than challenge with wild-type C. albicans. **P <0.01, *P < 0.05, compared with wild-type (WT) by one-way ANOVA, ns = not significant.