Xenbase Image ID: 169929

Figure 4. AhR controls Ccno expression and regulates the transcriptional programme required for ciliogenesis.(a,c) mRNA expression levels of the indicated genes in Ahr+/+ and Ahr−/− mTEC cultures at indicated days of ALI. Values are normalized to Hprt1. Mean±s.e.m.; two-way analysis of variance (variables: genotype and time). Data representative of at least three independent experiments. (b) Immunoblot and densitometric analysis of AhR protein expression as compared with β-actin, in AhR-sufficient and AhR-deficient mTEC cultures at days 0 and 8 of ALI. Data representative of three independent experiments. (d) Chromatin immunoprecipitation (ChIP) analysis of AhR interaction with the Ccno promoter in wt and Ahr−/− mTEC cultures at indicated days of ALI. Mean±s.e.m.; Student's t-test (unpaired, two-tailed). Data representative of three independent experiments. (e) Luciferase reporter assay was performed in AhR-sufficient and AhR-deficient mTEC cultures transfected at 5 days of ALI and analysed 36 h post transfection. Delta Ccno promoter lacks predicted AhR response elements. Mean±s.e.m.; Student's t-test. Data representative of two independent experiments. (f) Luciferase reporter assay was performed in NIH3T3 cells. Cells were co-transfected with a plasmid encoding AhR (pCMV-Ahr) or control vector (pCMV) and a luciferase-encoding reporter gene downstream of the Ccno promoter. Cells were analysed 36 h post transfection. Delta Ccno promoter lacks predicted AhR response elements. Empty indicates the control reporter vector. Mean±s.e.m. Student's t-test. Data representative of two independent experiments. (g) mRNA expression levels of Cdc20b (normalized to Hprt1) and mir449c (normalized to U6) in Ahr+/+ and Ahr−/− mTEC cultures at indicated days of ALI. Mean±s.e.m.; two-way analysis of variance (variables: genotype and time). Data are representative of three independent experiments. FC, fold change. Image published in: Villa M et al. (2016) Copyright © 2016, The Author(s). This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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