Figure 4.
AHR regulates Jund expression in lung epithelial cells. (A-C) Lethally irradiated B6.WT or B6.Ahr−/− mice received TCD BM cells and T cells from BALB/c mice. (A) Bulk RNA sequencing analysis of 5-day whole lungs. Heat map visualization of log2 (fold change; Ahr−/−/WT) is presented. (B-C) AP-1 family gene expression levels were measured in whole lungs (B) or isolated lung epithelial cells (C) by real-time polymerase chain reaction (PCR). (D) Isolated lung epithelial cells from B6.WT or B6.Ahr−/− mice were stimulated with IFN-γ for 48 hours, and IL-1β was added and further cultured for 24 hours. Jund and Fos expression levels were measured using real-time PCR. (E) Isolated lung epithelial cells from B6.WT mice were stimulated with IFN-γ in the presence or absence of tanshinone IIA (1 μM) for 48 hours, and IL-1β was added and further cultured for 24 hours. Il6 and Tnf expression levels were measured using real-time PCR. (F) A549 cells were cultured in the presence of L-Kyn (100 μM) with or without CH-223291 (10 μM) for 48 hours and further cultured in the presence of IL-1β for 24 hours. Levels of Jund and Il6 expression were measured by real-time PCR. (G) A549 cells were stimulated with IL-1β in the presence of the indicated inhibitors for 24 hours. Levels of Jund were measured by real-time PCR. (H) A549 cells were stimulated with IL-1β for 1 hour and further cultured in the presence of L-Kyn for 2 hours. Chromatin immunoprecipitation (ChIP) assays were performed to assess the binding of AHR to the xenobiotic response element site of the Jund promoter. (I-J) A549 cells were stimulated with IL-1β for 1 hour and further cultured in the presence of L-Kyn for 2 hours. ChIP assays were performed. (I) Binding of STAT1 (left) and JunD (right) to STAT1- and JunD-binding site of the Jund promoter, respectively. (J) Binding of JunD to a JunD-binding site of the Il6 promoter. Results are representative of 2 (A,I) or 3 independent experiments with similar results. Data represent mean ± standard error of the mean. Nonparametric Mann-Whitney U test (B-E) and 1-way analysis of variance (F-J) were performed. *P < .05, **P < .01, ***P < .001. mRNA, messenger RNA.

AHR regulates Jund expression in lung epithelial cells. (A-C) Lethally irradiated B6.WT or B6.Ahr−/− mice received TCD BM cells and T cells from BALB/c mice. (A) Bulk RNA sequencing analysis of 5-day whole lungs. Heat map visualization of log2 (fold change; Ahr−/−/WT) is presented. (B-C) AP-1 family gene expression levels were measured in whole lungs (B) or isolated lung epithelial cells (C) by real-time polymerase chain reaction (PCR). (D) Isolated lung epithelial cells from B6.WT or B6.Ahr−/− mice were stimulated with IFN-γ for 48 hours, and IL-1β was added and further cultured for 24 hours. Jund and Fos expression levels were measured using real-time PCR. (E) Isolated lung epithelial cells from B6.WT mice were stimulated with IFN-γ in the presence or absence of tanshinone IIA (1 μM) for 48 hours, and IL-1β was added and further cultured for 24 hours. Il6 and Tnf expression levels were measured using real-time PCR. (F) A549 cells were cultured in the presence of L-Kyn (100 μM) with or without CH-223291 (10 μM) for 48 hours and further cultured in the presence of IL-1β for 24 hours. Levels of Jund and Il6 expression were measured by real-time PCR. (G) A549 cells were stimulated with IL-1β in the presence of the indicated inhibitors for 24 hours. Levels of Jund were measured by real-time PCR. (H) A549 cells were stimulated with IL-1β for 1 hour and further cultured in the presence of L-Kyn for 2 hours. Chromatin immunoprecipitation (ChIP) assays were performed to assess the binding of AHR to the xenobiotic response element site of the Jund promoter. (I-J) A549 cells were stimulated with IL-1β for 1 hour and further cultured in the presence of L-Kyn for 2 hours. ChIP assays were performed. (I) Binding of STAT1 (left) and JunD (right) to STAT1- and JunD-binding site of the Jund promoter, respectively. (J) Binding of JunD to a JunD-binding site of the Il6 promoter. Results are representative of 2 (A,I) or 3 independent experiments with similar results. Data represent mean ± standard error of the mean. Nonparametric Mann-Whitney U test (B-E) and 1-way analysis of variance (F-J) were performed. *P < .05, **P < .01, ***P < .001. mRNA, messenger RNA.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal