Immunohistochemistry and Immunofluorescence of PLAU and LAMC2

HNSCC samples and normal samples were collected from humans for IHC; the methods for IHC are described in our previous study 19 (link). IHC was performed with anti-PLAU (17968-1-AP, Proteintech, 1:500) and anti-LAMC2 (ab210959, Abcam, 1:500) antibodies. The methods of immunofluorescence staining of PLAU and LAMC2 were similar to those of IHC. Sections were subjected to rehydration and antigen retrieval, followed by elimination of autofluorescence. Primary antibodies were then applied and incubated at 4°C overnight, followed by fluorescence-conjugated secondary antibody incubation and DAPI staining of nuclei.