Ab129202

The TMAs of the Zhongshan cohort were constructed as previously described.^{19 (link)} Dual immunohistochemistry (IHC) staining was performed. Briefly, the TMA slides were dewaxed in an oven and treated with water bath-heated xylene and graded alcohols. The slides were heated in sodium citrate buffer (0.01 M sodium citrate buffer, pH = 6) for 15 min for antigen retrieval. Normal goat serum blocking solution was applied for 20 min at 37 °C. Then, anti-CD103 antibody (Abcam, ab129202, diluted at 1:300) was applied for 2 h at 37 °C. After the primary antibody incubation, a general two-step kit detection system was used (HRP, Mo/Ra; ZSGB Biotech, PV-9000) with DAB. The slides were washed again and incubated with anti-CD8A primary antibody (Abcam, ab199016, diluted at 1:1500) overnight at 4 °C. TMA slides were subsequently washed, incubated with AP-labelled secondary antibody and stained with Vector Blue (Vector Blue AP Substrate Kit detection system; Vector Labs, SK-5300). Finally, the sections were washed, dehydrated and mounted. For immunofluorescence staining, the sections were incubated with the primary antibodies overnight at 4 °C. Then, samples were incubated with FITC-conjugated and TRITC-conjugated secondary antibodies for 2 h at 37 °C. Finally, the slides were mounted with anti-fade mounting solution containing DAPI. The slides were captured using a Leica DMi8 microscope.