Fig. 4: Psrc1 upregulates Ahr expression through attenuation of ubiquitination-mediated degradation in macrophages.

a Psrc1 mRNA and protein expression levels in were determined using RT–qPCR (top) and western blotting (bottom). b Ahr and Cyp1a1 mRNA expression levels were determined using RT–qPCR (n = 5). c Ahr protein expression was determined using western blotting. Two duplicate lanes were used as technical repetitions (n = 3). d Representative images of immunofluorescence staining of Ahr protein and DAPI for nuclei (n = 6). e Ahr protein expression in the cytoplasmic and nuclear fraction was determined using western blotting (n = 3). f After the treatment with CHX (15 μg/ml) for the indicated durations, Ahr protein expression was analyzed by western blotting. g Psrc1 protein expression levels were determined using western blotting. h BMDMs were treated with siCtrl or siPsrc1 for 48 h followed by DMSO or MG132 (10 μM) treatment for 6 h. Ahr protein expression levels were determined using western blotting (n = 3). i,j Uchl3 protein expression levels in BMDMs treated with AdPsrc1 (i) or siPsrc1 (j) were determined using western blotting (n = 3). k BMDMs treated with AdGFP or AdPsrc1 for 48 h followed by DMSO or TCID (10 μM) treatment for 24 h and MG132 treatment for 6 h before immunoprecipitation of Ahr and western blot of ubiquitin. Data are presented as mean ± s.d. Statistical analysis was performed by two-tailed unpaired t-tests (b, c, i and j), Mann–Whitney U test (a) and two-way ANOVA followed by Tukey’s post hoc test (h). BMDMs were treated with AdGFP or AdPsrc1 for 72 h in a–f and i. BMDMs were treated with siCtrl or siPsrc1 for 48 h in g, h and j.