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. Author manuscript; available in PMC: 2015 Nov 1.
Published in final edited form as: J Immunol. 2014 Sep 29;193(9):4643–4653. doi: 10.4049/jimmunol.1401625

FIGURE 4.

FIGURE 4

OGG1-BER-dependent expression of proinflammatory chemokines and cytokines. (A,B) Expression of proinflammatory mediators upon OGG1-initiated DNA base excision repair (A) and after 8-oxoG (B) challenge of OGG1-expressing (Cont-RNAi, left panel) or OGG1-deficient (Ogg1-RNAi, right panel) lungs. RNAs were isolated at from five lungs and pooled, and the synthesized cDNAs were analyzed by Mouse Inflammatory Cytokines & Receptors PCR Array (2h time point is shown). (C) Time course of OGG1-dependent expression of Cxcl1 (upper panel) and Cxcl2 (lower panel) mRNAs upon 8-oxoG challenge (n=5–6; □, Cont-RNAi; ■, Ogg1-RNAi). (D) KRAS-dependent expression of Cxcl1 and Cxcl2 after 8-oxoG challenge to the airways. KRas was depleted from the airways by RNAi. RNAs isolated after 8-oxoG challenge were analyzed by qRT-PCR (n=5–6, 2 h time point is shown). (E) Expression of CXCL1 in the airway epithelia (white arrow) of OGG1-proficient (Cont-RNAi) and OGG1-deficient airways (Ogg1-RNAi), as shown by IHC. Representative lung sections (upper panels) are shown after a 2h 8-oxoG challenge (DAPI-staining of sections is shown on lower panel). (F) Levels of CXCL1 in the BALF of mice (n=5) challenged with 8-oxoG (left panel) or in response to OGG1-BER (n=6) as determined by BioPlex assays. (G) KRAS-, but not NRAS-expression-dependent recruitment of neutrophils to airways. RAS was depleted by RNAi to Kras or Nras, and airways (n=5–6) were challenged with 8-oxoG. Neutrophil numbers were determined 16 h post-challenge. (H) OGG1-BER-induced recruitment of neutrophils to airways is KRAS-, but not NRAS-expression dependent. RAS were depleted in airways (n=5–6) as in the legend to G, and neutrophil numbers were determined 16 h post-challenge. (B,C,D,F,G) Lungs were challenged with 60 μL saline containing 0.0005 mg per kg 8-oxoG. (A,F,H) Oxidative DNA damage was generated by challenging the airways with glucose oxidase (Materials and Methods). *** = P<0.001). Cxcl1, chemokine (C-X-C motif) ligand 1 transcript; Cxcl2, chemokine (C-X-C motif) ligand 2 transcript; Ccl20, C-C motif chemokine 20; Ccl3, chemokine (C-C motif) ligand 3; Il1a, Interleukin-1 alpha; Il1b, Interleukin-1 beta; Tnfa, tumor necrosis factor; CXCL1, chemokine (C-X-C motif) ligand 1. *** = P<0.001; **** = P<0.0001.