Figure 10.

Rev-erbα inhibits CCL21 in gastric mucosa during H pylori infection. (A) The mRNA expression profiles of chemokines in gastric mucosa of WT H pylori–infected WT and Rev-erbα^–/– mice 8 weeks p.i. was analyzed by real-time PCR (n = 5). (B–D) The mRNA expression or the concentrations of CCL21 in (B) gastric mucosa of WT H pylori–infected WT and Rev-erbα^–/– mice, (C) gastric mucosa of WT H pylori–infected mice injected with Rev-erbα agonist SR9009 or cremophor control, or Rev-erbα antagonist SR8278 or DMSO control, or (D) gastric mucosa of WT H pylori–infected BM chimera mice were compared 8 weeks p.i. (n = 5). (E) The correlation between Rev-erbα expression and CCL21 expression in gastric mucosa of WT H pylori–infected WT mice 7, 8, and 9 weeks p.i. or in gastric mucosa of H pylori–infected patients was analyzed. (F) Representative immunofluorescence staining images showing CCL21-expressing (red) CD326⁺ cells (green) in gastric mucosa of H pylori–infected patients. Scale bars: 20 μm. (G) AGS cells were cotransfected with CCL21-luc construct or a mutant construct and Rev-erbα-pcDNA3.1 or pcDNA3.1 (control vector) for 48 hours. Luciferase activity was measured to assess CCL21 promoter activity (n = 3). (H) Representative data and statistical analysis of chromatin immunoprecipitation assay in AGS cells infected with WT H pylori or ΔcagA, followed by regular PCR with primers designed for Rev-erbα binding site of CCL21 promoter region (n = 3). (I) Primary GECs from uninfected Rev-erbα^–/– and WT mice, and Rev-erbα siRNA, NC, or lipo2000-only (Mock) pretreated AGS cells or AGS cells without treatment (medium) were infected with WT H pylori (MOI = 100) for 24 hours. CCL21 production was measured in cell culture supernatants by ELISA (n = 3). ∗∗P < .01, n.s. P > .05 for groups connected by horizontal lines.