Meso scale kits

Plasma EVs from above were lysed using M-PER™ Reagent (Thermo Scientific), supplemented with protease and phosphatase inhibitors (Roche). Equal volume of EV lysates were used to quantitatively measure proteins in the following Meso Scale kits (Meso Scale Diagnostics): total p53, phospho-p53 (Ser15), PARP (cleaved), and Caspase-3 (cleaved) by the Apoptosis Whole Cell Lysate Kit (Cat#K15102D-1); phospho-JNK (Thr183/Tyr185), phospho-p38 (Thr180/Tyr182), and phospho-ERK1/2 (Thr/Tyr: 202/204; 185/187) by the Apoptosis Whole Cell Lysate Kit (Cat# K15101D-1) and a custom made kit for phospho-Tau181 kit. Meso Scale kits were analyzed on a Meso Quickplex SQ 120. LC3 was measured using an ELISA kit (Cat# MB-S2602917 MyBioSource, Inc.). Clusterin was quantified by the Clusterin Competitive ELISA Kit (Cat# AG-45A-0013YEK-KI01 Adipogen Corporation).

Plasma levels of IFNβ, IFNγ, IL6, IL8, IL10, CXCL10, CXCL5, CCL20, TNFα, MCP1, PD-1, and PD-L1 were quantified by commercially available Meso Scale kits (Meso Scale Discovery, Rockville, MD, USA, cat# K15067, F214A-3 and F214C-3) in cryopreserved plasma samples from baseline and on-treatment samples according to manufacturer’s protocol. Data was acquired using a QuickPlex SQ 120 instrument (Meso Scale Discovery, Rockville, MD, USA). Samples below lower limit of quantification were set to zero.