Supplementary Figure 1: TEXs inhibit the innate antiviral response

(a, b) qPCR analysis of IFNB1 mRNA synthesis (a) and ELISA analysis of IFN-β secretion (b) by THP1 cells co-cultured with 16HBE and A549 cells for 24 h, followed by treatment with PBS, SeV or HSV-1 (MOI (multiplicity of infection), 10). (c) qPCR analysis of IFNB1 mRNA in THP1 cells co-cultured with or without control or Rab27a-depleted A549 cells (left) and in RAW264.7 cells co-cultured with or without control or Rab27a-depleted LLC cells (right) for 24 h, followed by treatment with PBS, SeV or HSV-1 (MOI, 10) for 8 h. (d) qPCR analysis of IFNB1 mRNA in THP1 cells co-cultured with DMA (25 μg ml⁻¹, 12 h)- pretreated A549 (left) or LLC (right) cells for 24 h, followed by infection with SeV or HSV-1 (MOI, 10) for 8 h. (e) qPCR analysis of IFNB1 mRNA in THP1 cells pretreated with control liposomes or MDA-MB-231-secreted exosomes (40 μg) for 24 h, followed by treatment with PBS, SeV or HSV-1 (MOI, 10) for 8 h. (f) Mass spectrometry identification of EGFR in MDA-MB-231 cells-secreted exosomes. (g) Immunoblot analysis of total cell lysate (TCL) and exosomes (Exo) derived from A549 and LLC cells. (h) Representative immunogold electron microscopy images of p-EGFR in exosomes derived from A549, LLC and lung cancer cells from two independent patients (P1 exo and P2 exo). (i) FACS analysis showing the percentage of EGFR⁺ exosomes secreted by A549 cells (n = 3 biological replicates). Negative control: IgG. (j) Immunoblot analysis of cell lysates from THP-1, bone marrow-derived macrophages (BMDM) and primary peritoneal macrophages (PM) treated with 16HBE-, A549-, MLF- and LLC-secreted exosomes (exo, 40 μg, 24 h) as indicated. (k) FACS analysis with human EGFR specific antibody showing the percentage of EGFR⁺ RAW 264.7 cells after pretreatment with A549 secreted exosomes (exo) (n = 3 biological replicates). Negative control: IgG. All qPCR results are shown as mean + s.d. of triplicates of at least two independent experiments. *P < 0.05 (two-tailed Student’s t-test (a–e).