Immunostaining Protocols for Cell and Tissue Analysis

Immunocytochemistry (ICC) was essentially as described (19 (link)). Primary antibodies as noted above CK5 (1:100), CK17 (1:200); vimentin (1:200), or CK8/18 (1:2000) for 2 h, secondary antibodies (A11029, A11037, A11008, A11032, Invitrogen, 1:200) for 1 h, and counterstained with DAPI. Cells were imaged using the Olympus BX40 fluorescent microscope and channels merged in Adobe Photoshop 2021. Immunohistochemistry (IHC) was performed as previously described (33 (link)) using CK5 (rabbit, ab75869, Abcam, 1:200), CK17 (rabbit, ab109725, Abcam, 1:100), vimentin (mouse, 5G3F10, Cell Signaling Technologies, 1:100), or pan-CK (mouse ab86734, 1:400, abcam, Waltham, MA) antibodies and either fluorescent secondary antibodies as described and imaged using an Olympus BX40 fluorescent microscope, or ImmPRESS Peroxidase detection kit (Vector Laboratories).