Fig. 3

Integration of scRNA-seq and spatial transcriptomics reveals the spatial distribution of macrophages in HCC and functional validation of M2 polarization. (A-G) Spatial transcriptomics (ST) of HCC tissue sections showing the overall transcriptional landscape. (H-N) Single-cell RNA sequencing (scRNA-seq)-derived cell type annotations mapped onto the ST data, identifying major cell types. (O) Transwell co-culture model illustrating the interaction between tumor cells (upper chamber) and macrophages (lower chamber). (P) Quantitative RT-PCR analysis comparing the expression of M2-specific marker genes in macrophages co-cultured with tumor cells versus macrophages cultured alone. (Q) Western blot analysis showing the protein levels of CD206, iNOS, ARG-1 in macrophages after co-culture with tumor cells compared to controls and quantifications (n = 3). (R) ELISA analysis of the culture medium for the secretion level of TNF-α, IL-1β, TGF-β, and IL-6 (n = 6).