Fig. 6

Synergistic effect of inhibiting DDR1 with nilotinib on aPD-1 in cirrhotic HCC through reversing neutrophils/NETs-dominant immunosuppressive TME. A Level change of phosphorating DDR1 in Col1-treated HepG2 cells w/o nilotinib and DDR-IN-1 by Western Blot. B CXCL8 mRNA level change in Col1-treated HepG2 w/o nilotinib by qPCR. C Neutrophils chemotaxis to Col1-treated HepG2 cells w/o nilotinib by flow cytometry in Transwell system. D–F Tumor growth (D), IHC images (E) and flow cytometry quantification (F) of infiltrated neutrophils in Hepa1-6 subcutaneous mice model challenged with cirrhotic-ECM plus nilotinib (nilo)/aPD-1/nilo+aPD-1. Scale Bar: 100 μm. G–M Experiment design was shown as G. Representative images (H) and quantification (I) of tumor growth, IHC/IF staining (J) and quantification of Ly6G+ neutrophils (K), CD8+ T cells (L) and NETs (M) in Hepa1-6 orthotopic HCC mice model with DMN induced cirrhosis treated with nilotinib (nilo)/aPD-1/nilo+aPD-1. Scale Bar: 100 μm. Cell number was calculated by counts per ×40 view. N A schematic diagram described the mechanism of cirrhosis ECM induced NETs performed blockade of T cell cytotoxicity and attenuated ICI efficiency in HCC. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001