Fig. 1

Opal six-colour multiplex immunofluorescence images of HCC tissues and levels of different immune targets in HCC. a Representative mIF and H&E staining images of HCC tissues. Arrow in case 1, Cytokeratin18⁺FGL1⁺ cells. Arrow in case 2, Cytokeratin18⁺PD-L1⁺ cells. DAPI was used to visualize nucleus (blue). Distribution of levels of LAG-3⁺ cells (b), CD8⁺ T cells (c), PD-L1 on TC (d), PD-L1 on IC (e), FGL1 on TC (f) and FGL1 on IC (g) in 143 HCC cases. The frequency of positive or high expression for each marker was displayed in parenthesis. The detailed cut-points used to define positive or high expression of these markers were described in methods. N negative, P positive, TC tumor cells, IC immune cells