ISG20 promotes local tumor immunity and contributes to poor survival in human glioma

Gao, Mengqi; Lin, Yi; Liu, Xing; Li, Yiming; Zhang, Chuanbao; Wang, Zheng; Wang, Zhiliang; Wang, Yulin; Guo, Zongze

Immunohistochemistry was performed using formalin-fixed, paraffin-embedded tissues. Four μm-thick sections were cut and dewaxed in xylene, rinsed in graded ethanol, and rehydrated in distilled water. After antigen retrieval with sodium citrate buffer (10 mM sodium citrate, pH 6.0), endogenous peroxidase activity was blocked with 3% H 2 O 2 . Then the sections were added with the appropriate antibody at appropriate dilution, then DAB staining solution was added. The sections were counter-stained with hematoxylin, dehydrated, and sealed. For immunofluorescence, antigen was retrieved with EDTA buffer (1 mM Tris/EDTA, pH 9.0) and endogenous fluorescence was eliminated with AutoFluo Quencher (Servicebio, Cat# G1221). The details of the antibodies used for immunofluorescence and immunohistochemistry are given in Table S6 . Staining for each marker was scored using Image-pro plus (v.6.0). Integrated optical density (IOD) to area ratio was calculated for each marker to assess the staining intensity.