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. 2024 Jun 4;116(10):1598–1611. doi: 10.1093/jnci/djae120

Figure 6.

Figure 6.

Batf knockout decreased interleukin 23 (IL-23)-IL-23R signaling in Pten-deficient mice, and Batf binds to the motifs of the mouse IL-23R gene promoter region. A) Representative immunohistochemistry for IL-23R–positive immune cells of Batf+ and Batf- mice stroma at different ages. Original magnification, ×100 (scale bar, 200 µm); inert, ×400 (scale bar, 50 µm). B) Numbers of IL-23R–positive cells per high-power field. Data are mean (95% confidence interval), n = 3 mice/group. *P < .05, **P < .01. C) Representative Western blot result for IL-23R in prostate tissue of Batf+ and Batf- mice at 9 and 30 weeks of age. D) Quantitative reverse transcriptase–polymerase chain reaction (PCR) results of Il23r in the prostate tissue of 12-week-old Pten wild-type mice (Ctr) and Pten-null mice. E) Quantitative reverse transcriptase–PCR results of Il23r in the prostate tissue of 12-week-old Batf+ and Batf- (Pen-null), and Batf knockout (Pten wild-type) mice. F) Quantitative reverse transcriptase–PCR results of Il23r in the prostate tissue of 30-week-old Batf+ and Batf- mice. G) Quantitative reverse transcriptase–PCR results of Il23p19 in the prostate tissue of 12-week-old Batf+ and Batf- (Pten-null), and Batf knockout (Pten wild-type) mice. H) Quantitative reverse transcriptase–PCR results of Il23p19 in the prostate tissue of 30-week-old Batf+ and Batf- mice. I) Mouse IL-23R levels in the plasma of Batf+ and Batf- mice. J) Representative Western blot for IL-23R in CD4+ T cell lysates from Batf+ and Batf- mice. K-L) Quantitative reverse transcriptase–PCR results for Il23r messenger RNA in Batf+ and Batf- mouse splenocytes (K) or in ex vivo cultured mouse prostate tissue (L) in Th17 differentiation conditioned media for 72 hours. *P < .05, **P < .01, ***P < .001. M) IL-23R binding sites and chromatin immunoprecipitation (ChIP) quantitative PCR product information. N, O) The results of ChIP assays were measured by quantitative PCR. Enrichment of IL23R binding sites using anti-Batf rabbit polyclonal antibody on sheared chromatin from mouse splenocytes. Normal rabbit immunoglobulin G (IgG) was used as a negative immunoprecipitation (IP) control. The purified DNA was analyzed on the Bio-Rad CFX Opus 96 Real-Time PCR System, with optimized primers for the promoter region of the AP-1 motifs in the IL23R gene. Data are presented as fold enrichment of the antibody signal vs the negative control IgG without the IL-23 treatment group, calculated using the comparative Ct method (also referred to as the 2-△△Ct method). Data are mean (95% confidence interval) (n = 3) of 3 independent experiments. *P < .05, **P < .01, ns = not significant difference compared with the corresponding IgG IP groups.