Mediators of Inflammation / 2019 / Article / Fig 3

Research Article

PARP-1 Is Critical for Recruitment of Dendritic Cells to the Lung in a Mouse Model of Asthma but Dispensable for Their Differentiation and Function

Figure 3

WT or PARP-1−/− mice were subjected to OVA sensitization followed by a single challenge or left unchallenged. A group of mice received olaparib (5 mg/kg) 30 minutes post-OVA challenge. Mice were sacrificed 48 h later. Lungs from the different experimental groups were fixed with formalin or processed to generate single-cell suspensions. (a) Lung sections were stained with hematoxylin and eosin; bar: 50 μm. (b) Cells were stained with a combination of antibodies to CD45, CD11b, and CD11c. CD11b+/CD11c+ cell population was gated from the live CD45+ population. (c) WT or PARP-1−/− mice were sensitized twice with OVA; spleens and lymph nodes were then collected 6 hours after the last sensitization and processed for single-cell suspensions. Cells were then stained with a combination of antibodies to CD11b, CD11c, and MHC. (d) Percentage of CD11b+/CD11c+/MHCIIhigh DCs that express CD80, CD86, or CD40. (e) Sorted CD11c+/CD11b+/MHCIIhigh cells from OVA-sensitized WT or PARP-1−/− mice were pulsed with OVA 323-339 peptide or vehicle overnight, washed, and then cocultured with CFSE-stained CD4+ T cells from OTII mice for four days. Proliferation was assessed by flow cytometry. For (a, b) and (d–h), the results are expressed as ; ; ; .
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