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. 2023 Nov 8;8:64. doi: 10.1038/s41536-023-00339-7

Fig. 4. AHR transcriptionally increased IL-22RA1 expression in mTEC1 cells.

Fig. 4

a mTEC1 cells were treated by vehicle or colivelin (1 μM) for 24 h. Immunoprecipitation was performed with anti-STAT3 or anti-AHR (n = 3). Western blot was used to detect proteins in the precipitate. b mTEC1 cells were treated by vehicle, stattic (1 μM) or CH-223191 (10 μM) for 24 h. ChIP assay was performed with anti-STAT3 as described (n = 5). c, d mTEC1 cells were treated by vehicle, stattic or colivelin for 24 h with/without FICZ (100 nM). qPCR was performed to detect mRNA levels in cells as described (n = 5). e mTEC1 cells were treated by colivelin and/or FICZ for 24 h (n = 5). Luciferase reporter assay was performed as described. f, g Total proteins were extracted from thymic stromal cells isolated from irradiated wild type mice which were treated by vehicle or FICZ (n = 3). Whole-cell proteins were extracted from mTEC1 cells treated by vehicle, FICZ (100 nM) or colivelin (1 μM) for 24 h (n = 3). Western blot was used to detect proteins as indicated. h Wild type C57BL/6 mice, pretreated by 5.5 Gy TBI, were intraperitoneally injected with vehicle, FICZ (0.1 mg/kg) or combined FICZ (0.1 mg/kg) and stattic (10 mg/kg) thrice weekly for 2 weeks. At day 14, thymus cells were counted as described (n = 5). i mTEC1 cells were treated by vehicle or rmIL-22 (20 ng/ml) in the presence of different doses of CH-223191 for 24 h (n = 5). Cell viability was detected by CCK-8 assay. Data are mean ± SD, compared using one-way ANOVA test or Student’s t test. *, p < 0.05; **, p < 0.01; ***, p < 0.001; n.s., not significant.