Fig. 3

Effects of IFN-γ on sEVs derived by SACC-83. a Representative TEM images of the purified control sEVs and IFN-γ-induced sEVs secreted by SACC-83. Scale bar: 50 nm. b Western blot analysis indicating the presence of the EV makers CD9, Alix and Tsg101, and the absence of the negative marker GM130 in the purified sEVs. c Characterization of the purified sEVs by NTA. d Comparison of the sizes between control sEVs (C) and IFN-γ-induced sEVs (IFN-γ) derived by SACC-83 (n = 4). e Curves of the sEV secretion of SACC-83 cells upon IFN-γ treatment (n = 3). f Comparison of the numbers of sEVs between control group (C) and IFN-γ treatment group (IFN-γ) derived by SACC-83 (n = 4). g, h Comparison of the protein contents of sEVs between control group (C) and IFN-γ treatment group (IFN-γ) (n = 4). i Western blot analysis of Rab27a and Hrs in the control cells and IFN-γ treated cells (IFN-γ). j Representative contour plots of the control sEVs and IFN-γ-induced sEVs. k–m Comparisons of the ratios of PD-L1⁺, CD54⁺ and PD-L1⁺CD54⁺ sEVs between control sEVs (C) and IFN-γ-induced sEVs (IFN-γ) respectively (n = 3). n Diagram of quantification of sEV PD-L1 by ELISA. o sEV PD-L1 were measured by ELISA (n = 3). p Western blot analysis of CD54 and PD-L1 in the control sEVs and IFN-γ-induced sEVs. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.000 1