In summary, we tested the expression pattern(s) of several positive (SMA, S100A4, TE-7, CD90, FAP-A, and PDGFRA) and negative (EpCAM, CK8, 18, CD45, and CD31) CAF markers and markers associated with CAF functions (TGFB1, TGFB2, TGFR1, TGFR2, and CD44), as well as immune markers (PD-L1, PD-L2, and PD-1) in the primary CAFs as cultured from resected tumor samples of two patients using four methods, qRT-PCR (Figure 3), flow cytometry (Figure 4A), Western blot (Figure 4B), and ICC (Figure 5). The gene discussed is TGFB1; the disease is intrahepatic cholangiocarcinoma.