Figure 2.

S. aureus clearing and survival of infected BMDM depend on functional NOX2. BMDM were infected with GFP-expressing S. aureus MW2 (MOI15) opsonized with 5% NMS in suspension and assessed 24 h p.i. by flow cytometry for clearing of S. aureus and for BMDM viability (right panel). (A) BMDM pre-treated with 10 μM DPI (open circles) or left untreated (closed circles), (B) wildtype (closed circles) and nox2^−/− BMDM (open circles), (C) wildtype (closed circles) and nox1^−/− BMDM (open circles). Statistical significance was analyzed by unpaired two-sided Student's t-test using GraphPad Prism 5.01 between indicated conditions (^** = p < 0.01; ^***= p < 0.001). The viabilities of BMDMs at 24 h p.i. were assessed by trypan blue exclusion and depicted as bar charts (A–C, right). Represented is the percentage of viable cells referring to corresponding non-infected cells for each genotype in mean and SD of triplicates. Statistical significance was analyzed by 1-way ANOVA with Bonferroni post-test between selected pairs as implemented in GraphPad Prism 5.01 between indicated conditions (^*= p < 0.05; ^** = p < 0.01; ^***= p < 0.001). (D) Clearing kinetics of wildtype, p22phox^Y121H and nox4^−/− at 6h post infection. Shown are means and SD of triplicates as percent of corresponding values at t0. Shown are representative experiments performed three times. NOX, NADPH Oxidas; ROS, reactive oxygen species; PMA, phorbol 12-myristate 13-acetate; DPI, diphenyleniodonium; MOI, multiplicity of infection; BMDMs, bone marrow derived macrophages; SD, standard deviation; TLR2, Toll-like receptor 2; NMS, normal mouse serum; p.i., post infection; CFU, colony forming unit; i.v., intravenous; s.c., subcutaneous.