Expression level of PCDH8 was determined based on the immunohistochemistry protocol (Paraffin) from Cell Signaling Technology (Beverly, MA) as previously described.21 (link) Immunohistochemical analysis was performed using PCDH8 antibodies (1:100 dilution, Abcam, Cambridge, MA) and secondary antibody conjugated with HRP (SuperPicture Polymer Detection Kit, HRP, broad spectrum; Thermo Fisher Scientific, Inc., Waltham, MA).
Assessment of PCDH8 staining was conducted according to staining showing and positive cells percentage. Thereby, PCDH8 staining patterns were categorized into 5 groups: 1+, less than 10% of cells stained positive; 2+, 10% to 25% positive cells; 3+, 26% to 50% positive cells; 4+, 51% to 75% positive cells; 5+, greater than 75% positive cells. The PCDH8 staining images for IHC and hematoxylin-eosin staining were obtained using the NanoZoomer 2.0-RS system (Hamamatsu Photonics Inc., Germany), and the digital slides were analyzed by NDP. view 2.5.14 version.
Assessment of PCDH8 staining was conducted according to staining showing and positive cells percentage. Thereby, PCDH8 staining patterns were categorized into 5 groups: 1+, less than 10% of cells stained positive; 2+, 10% to 25% positive cells; 3+, 26% to 50% positive cells; 4+, 51% to 75% positive cells; 5+, greater than 75% positive cells. The PCDH8 staining images for IHC and hematoxylin-eosin staining were obtained using the NanoZoomer 2.0-RS system (Hamamatsu Photonics Inc., Germany), and the digital slides were analyzed by NDP. view 2.5.14 version.
