IHC
IHC was performed on 3-μm sections cut from paraffin blocks using fully automated systems. The CYP11B1 primary antibody clone RAT-87 (MABS502; Merck) was used at a dilution of 1:100 after heat-induced epitope retrieval at pH 9.0 (BOND Epitope Retrieval Solution 2; AR9640; Leica) for 20 min. CYP11B2 primary antibody clone EPR10494 (ab168388; Abcam) was used at a dilution of 1:200 after proteolytic-induced epitope retrieval with proteinase K (BOND Enzyme Pre-Treatment Kit; AR9551; Leica) for 10 min. The primary antibody binding to tissue sections was visualized using a BOND Polymer Refine Detection system (DS9800; Leica). IHC slides were scanned using a NanoZoomer S210 (C13239; Hamamatsu) and viewed using the Hamamatsu NDP.view2 image-viewing software (U12388-01; Hamamatsu).