Bip chop

Flow cytometric analysis of PLCs and hLSECs was performed using antibodies for TLR 3,4,7,8,9 (ab45093, ab11227, ab45097, ab58864, Abcam, Cambridge, UK [TLR 3,4,8,9] and IC5875P, R&D Systems [TLR 7]) and BiP/CHOP (Cell Signaling Technology, Danvers, MA). Intracellular staining was performed as per manufacturer’s protocol using the Intracellular Flow Cytometry kit (Cell Signaling Technology, Danvers, MA) for TLR 3,7,8,9 and BiP/CHOP. Cells were directly stained and fixed for TLR 4. After processing, cells were analyzed using a BD Accuri C6 flow cytometer, and the data were analyzed using FlowJo Software (BD Biosciences, San Jose, CA). Cells treated with tunicamycin (1 μg/mL overnight for BiP and 2 μg/mL 8 h for CHOP) were used as positive control for ER stress.