Anti ahr antibody

Cells were lysed on ice for 10‒20 min in RIPA buffer (Gibco, Carlsbad, USA). Cell lysates were then centrifuged (12,000
g), and supernatants were collected for western blot analysis. Equal amounts were separated by SDS-PAGE and blotted onto PVDF membranes (Millipore, Billerica, USA). The membranes were initially incubated with primary antibodies including anti-AHR antibody (Abcam, Cambridge, UK), anti-CYP1A1 antibody (Abcam), and anti-CYP1B1 antibody (Abcam), subsequently incubated with the corresponding secondary antibodies for 1 h. Specific bands were visualized using an Enhanced chemiluminescence (ECL) western blotting detection kit (Amersham Biosciences, Buckinghamshire, UK).

Pathological sections of prostatic tissues were retrieved using microwave antigen retrieval (Thermo, USA). Then samples were blocked by 5% Bovine Serum Albumin, followed by incubating with anti-TDO2 antibody (1:200, abcam, UK), anti-Kyn antibody (1:300, abcam, UK), anti-AhR antibody (1:200, abcam, UK), anti- NF-κB (1:200, abcam, UK) and anti-c-Myc antibody (1:500, abcam, UK) for 4 °C overnight. For immunofluorescence staining, the sections were then incubated with secondary antibodies (1:1000; abcam, UK), and the nucleus was stained with DAPI. Images were captured using a FV1000 laser scanning confocal microscope (Leica, Germany). For immunohistochemical staining, the sections were then stained using the ABC HRP Kit (Thermo, USA) and counterstaining with hematoxylin. Immunohistochemical images were captured using microscope (Leica, Germany). The intensity of proteins in sections were analyzed by Image-Pro Plus 2.0 software.