Immunofluorescence and Immunohistochemistry for FoxM1 and Ki-67 in Tissue Sections

Fixed and paraffin tissue sections were processed as described before (31 (link), 32 (link)). FoxM1 and Ki-67 (cat # 12202S Cell Signaling Technology, Inc., MA, USA) antibodies were used for immunofluorescence (IF) co-staining. For immunohistochemistry, secondary antibodies were applied, followed by incubation with Streptavidin-HRP conjugate (Invitrogen, NY, USA) and slides were developed with DAB (Vector Laboratories) and counterstained with Harris Haematoxylin, dehydrated, and mounted with Permount (Fisher Scientific, NJ, USA). For IF, goat anti-Rabbit IgG fluor and Goat anti-mouse IgG Texas Red (Invitrogen, NY, USA), secondary antibodies were applied, stained with DAPI, and mounted with Flour-G (Invitrogen, NY, USA). All images were captured using a Nikon 80i upright microscope (Tokyo, Japan) in the KUMC imaging center.