IL-32 is secreted from myeloma cells on EVs. (A) JJN3 cells were stained for IL-32 (red), GM130 (Golgi marker; green), CD63 (EV marker; green), and CD81 (EV marker; green). Scale bar, 15 μm. (B) Total cell lysates and lysates of EVs isolated from cell culture supernatants were immunoblotted as indicated. (C) EVs were isolated from media from JJN3 cells cultured for 48 hours. IL-32 in the EV fraction treated with or without NP-40 detergent was quantified by ELISA. (D) EVs were isolated from BM plasma obtained from patients with myeloma (n = 18), and amount of IL-32 in the EV fraction was determined by ELISA.