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. 2015 May 4;212(5):729–742. doi: 10.1084/jem.20141675

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© 2015 Bourdonnay et al.

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Figure 4. — SOCS1 protein is secreted in Exos and exerts inhibitory effects on AEC STAT1 activation. (A) Overnight AM CM (+) or RPMI 1640 alone (−) was concentrated and subjected to WB analysis for SOCS1; bar graph depicts arbitrary densitometric units of SOCS1. (B) MPs and Exos were isolated from overnight CM and subjected to WB for SOCS1. (C) AECs were pretreated for 2 h with (+) Exos isolated from overnight CM or with RPMI 1640 alone (−) before a 1-h stimulation with IFNγ, after which AEC lysate proteins were subjected to immunoblot analysis for p-STAT1. STAT1 activation was expressed as the percentage of p-STAT1, normalized for β-actin, in cytokine-treated AECs not pretreated with AM-derived Exos. (A–C) Data represent the mean ± SE from at least three independent experiments (A and C) or are representative of two independent experiments (B). *, P < 0.05 versus RPMI 1640 alone (A) or IFNγ-treated AECs not treated with Exos (C).