Ship 1

Total proteins were extracted from plasma exosomes using radioimmunoprecipitation (RIPA) lysis buffer (BioSharp, Hefei, China), and the concentration was determined using the Pierce bicinchoninic acid (BCA) protein assay kit (Thermo Fisher Scientific, Rockford, IL, USA). Equal amounts of each protein sample were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene fluoride (PVDF) membrane before incubation overnight with primary antibodies against CD63 and CD9 (1:1,500; Abcam, Cambridge, MA, USA), SHIP1, CDKN1B, and SOCS1 (1:800; Santa Cruz Biotechnology, Dallas, TX, USA), and β-actin and phosphorylated NF-κB p65 (1:1,000; Cell Signaling Technology, Beverly, MA, USA). Membranes were then incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies (1:4,000; Cell Signaling Technology). The protein bands were visualized using IMAGISOLANE LAS4000 Mini (GE Healthcare, Piscataway, NJ, USA). Image-Pro Plus 6.0 software (Media-controlnetics, Silver-Spring, MD, USA) was used to measure the scale values of protein bands, and the relative expression of the indicated proteins was normalized to the internal control β-actin.

Cells were lysed with lysis buffer (0.5% Nonidet P-40, 50 mM Tris-HCl (pH 8.0), 120 mM NaCl, 50 mM NaF, plus 1 mM Na₃VO₄, 10 g/ml aprotinin, 100 g/ml leupeptin and 10 mM phenylmethylsulfonyl fluoride). Lysates (40 μg) were fractionated by SDS/PAGE and transferred to a polyvinylidene fluoride membrane (Immobilon-P, Millipore, Billerica, MA, USA). The following antibodies were used: SHIP-1, Fli-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); β-actin (Sigma-Aldrich); goat-anti-mouse, and goat anti-rabbit HRP-conjugated (Promega, Madison, WI, USA), ERK, phospho-AKT, AKT, MYC and JAK2 antibodies from Cell Signaling Technology (CST, Danvers, MA, USA).