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

Fig. 3. Targeting AHR had an impact on IL-22RA1 expression in mTEC1 cells.

Fig. 3

a CH-223191 was used to treat mTEC1 cells for 24 h. b, c mTEC1 cells, stably transduced with control shRNA or Ahr-shRNA lentivirus, were treated with vehicle or FICZ (100 nM) for 24 h. d Total cell proteins were extracted from the mTEC1 cells as described above. Western blot was performed with indicated antibodies (n = 3). e FICZ was used to treat mTEC1 cells for 24 h. qPCR was performed to detect mRNA levels in cells as described (n = 5). f mTEC1 cells were treated by vehicle, CH-223191 (10 μM) or FICZ (100 nM) for 24 h. ChIP assay was performed with anti-AHR. The precipitated DNA was detected by qPCR with primers of mouse IL-22RA1 gene promotor (n = 5). g mTEC1 cells stably transduced with control shRNA or Ahr-shRNA lentivirus, h mTEC1 cells stably transduced with control or Ahr-overexpression lentivirus, i mTEC1 cells, without genetic modification, were treated by vehicle, CH-223191 (10 μM) or FICZ (100 nM) for 24 h. g, h, i These cells were transfected with pGL4.10 plasmids (firefly, following mouse IL-22RA1 gene promoter sequence) and pRL-TK plasmids (renilla, normalizer) for 12 h. Luciferase activity was measured (n = 5). 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.