Figure 1

Flowchart for patient classification and quantitative analysis of CTLs, CD4⁺CD25⁺ cells, granzyme B-expressing cells, and Foxp3-expressing cells in HCC tissues in this study. By NGS-based pre-S genotyping, HBV-related HCC patients were divided into two groups, the pre-S2 mutant-positive and -negative groups, according to either the presence of deletion spanning pre-S2 gene segment or the percentage of pre-S2 plus pre-S1 + pre-S2 deletion. By fluorescent IHC staining, CTLs, CD4⁺CD25⁺ cells, granzyme B-expressing cells, and Foxp3-expressing cells in HCC tissues were detected and quantified as the density of CTLs, CD4⁺CD25⁺ cells, granzyme B-expressing cells, and Foxp3-expressing cells for comparative analysis between the two groups of patients.