Tissue Microarray Analysis of TG2 Expression

Lung tissues were fixed in 4% paraformaldehyde, processed, and embedded in paraffin. We carefully examined the cores and inserted them into new paraffin blocks, using Tissue Arrayer Minicore (Alphelus, Plaisir, France). We deparaffinized the sections with a thickness of 5 μm and washed them with 100% ethanol, 90% ethanol, 70% ethanol, and then distilled water. These sections were then prepared for antigen retrieval in a citrate buffer with a pH of 6.0 by heating them in a microwave for 5-min cycles. The sections were then incubated overnight at a temperature of 4°C with a 1:200 diluted Anti-TG2 antibody (Abcam, USA), after which we incubated them with biotin-labeled Rabbit Anti-Mouse IgG H&L preabsorbed (Abcam, USA) for immunostaining (Chihong, et al., 2017 (link)).