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. 2018 Nov 15;198(10):1288–1301. doi: 10.1164/rccm.201712-2388OC

Figure 7.

Figure 7.

Rac2 (Ras-related C3 botulinum toxin substrate 2) ameliorates cigarette smoke (CS)–mediated lung infection. Wild-type mice were administered pLVX-IRES-tdTomato (empty) or pLVX-IRES-tdTomato-Rac2CA (Rac2CA) lentivirus. (A) Representative confocal images of CD11c-, Ly6c-, and RFP-positive BAL cells counterstained with DAPI. Scale bar, 25 μm (n = 3). Wild-type mice were transduced with empty or Rac2CA lentivirus. After 6 weeks, mice were exposed to air or CS and then infected with saline or Streptococcus pneumoniae. (B) Immunoblot analysis of isolated BAL cell membrane. (C) Immunoblot analysis of isolated BAL cell membrane and cytosol with corresponding (D) Rac2 activity in BAL cell membrane (n = 4–8). (E) NOX2 (NADPH oxidase 2)-derived reactive oxygen species generation in BAL cell membrane (n = 5). (F) Lung colony-forming units (cfu; n = 5). Hematoxylin and eosin staining was performed in (G) empty-transduced (n = 5) and (H) Rac2CA-transduced wild-type mice (n = 5). Scale bars, 600 μm. The second and fourth rows in G and H show magnifications of the boxed regions in the first and third rows, respectively. *P < 0.05; ***P < 0.0001. One-way ANOVA with Tukey statistical analysis were used for D–F. Representative immunoblots and histology micrographs are shown. Mem = membrane; OD = optical density; RFP = red fluorescent protein; S.p. = Streptococcus pneumoniae.