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Representative examples of immunostaining for CD3, CD8, CD163, PD-L1, and FOXP3 in patient #3 pre- and post-NACT in BTC. Pictures demonstrate an increase in the expression of CD3, CD8, CD163 and a decrease in PD-L1, FOXP3 post-NACT at 40× magnification using MICSSS, refer to methods section for details. TIL, tumor infiltrating lymphocyte; NACT, neoadjuvant chemotherapy; BTC, biliary tract cancer; MICSSS, multiplexed immunohistochemical consecutive staining on single slide.
