Fig. 4

Regulatory functions of LFNPs/siRNA complexes in vitro. A Cytotoxicity of tested complexes in 4T1 cells and exhausted T cells (n = 5). B Cellular uptake of FEGCG complexes and different LFNPs3 complexes detected by flow cytometry in 4T1 cells and exhausted T cells (n = 5). C Cellular uptake and lysosomal escape of FEGCG complexes and LFNPs3-3 complexes detected by confocal microscopy. Scale bar = 10 µm. D Luc gene silencing of FEGCG complexes and different LFNPs3 complexes with or without FBS (n = 5). E TOX expression detected by flow cytometry and western blotting after different treatments in exhausted T cells (n = 5). F PD-L1 expression detected by flow cytometry and western blotting after different treatments in 4T1 cells (n = 5). G Percentage of PD-1 + Tim-3 + cells detected by flow cytometry after different treatments in exhausted T cells (n = 5). *P < 0.05, **P < 0.01, ****P < 0.0001 by one-way analysis of variance (ANOVA) with Turkey’s multiple comparisons