FIGURE 3.
Generation of SPCCre/CARMA3F/F mice. (A) Generation of CARMA3 targeting construct. (B) Southern blot analysis to identify correctly targeted ES cell clones obtained from ES cells electroporated with the targeting construct. Two ES cell clones (C1 and C2) were identified in which there was the expected 5′ and 3′ recombinations. W, wild-type mouse. Immunohistochemistry of lungs from naive (C) SPCCre/CARMA3+/+, (D) SPCCre/CARMA3F/F, and (E) CARMA3F/F mice stained with an Ab against CARMA3 (top panels) or an isotype control Ab (bottom panels). Scale bars, 200 μm. (F) Immunofluorescence of lungs from naive SPCCre/CARMA3+/+ and SPCCre/CARMA3F/F mice stained with an Abs against CARMA3 and E-cadherin. All images were taken using the same exposure. Scale bars, 20 μm. Basal, ciliated, and secretory cells sorted from the lungs of SPCCre/CARMA3+/+ and SPCCre/CARMA3F/F mice that were either (G) naive or (H) received one OVA/alum immunization and one OVA challenge. RNA levels of CARMA3 were determined by qPCR. Data are means ± SEMs of six mice per group. *p < 0.05 (SPCCre/CARMA3+/+ compared with SPCCre/CARMA3F/F). (I) Immunofluorescence of lungs from PBS- and OVA-immunized and challenged SPCCre/CARMA3+/+ mice stained for CARMA3. Scale bars, 20 μm.