Microscope 1x701

Five-μm fixed pancreas sections from control and treated mice were incubated with primary antibodies in a hybridization chamber for 1 h at room temperature or overnight at 4°C. The primary antibodies used were mouse-specific PCNA (Cat#SC-56, Lot#I1212), caspase3, Bcl2, and p21 (Santa Cruz, CA). Following incubation with the primary antibody, sections were incubated for 1 h with anti-mouse or anti-rabbit secondary antibodies, as appropriate for each primary antibody, then were visualized with diaminobenzidine (DAB) and counterstained with hematoxylin for IHC. Slides were observed under an Olympus microscope 1X701 and digital computer images were recorded with an Olympus DP70 camera.

5-μm fixed sections were incubated with primary antibodies in a hybridization chamber for 1 h at room temperature or overnight at 4 °C. The primary antibodies for proliferating cell nuclear antigen (PCNA), Cav-1, β-catenin and p21 were from Santa Cruz Biotechnology ); those for Ki67 and cyclin D1 were from Cell Signaling; those for p16 were from LS Bio and those for Rb were from Abcam. Following incubations with primary antibody, sections were incubated for 1 h with anti-mouse or anti-rabbit secondary antibodies, as appropriate for each primary, then visualized with diaminobenzidine (DAB) and counterstained with Hematoxylin for IHC and counterstained with DAPI (4′,6-diamidino-2-phenylindole) for immunohistofluorescence (IHF). Slides were observed under an Olympus microscope 1X701 and digital computer images were recorded with an Olympus DP70 camera.