Journal: PLoS Pathogens
Article Title: Interferon regulatory factor 8 regulates caspase-1 expression to facilitate Epstein-Barr virus reactivation in response to B cell receptor stimulation and chemical induction
Figure Lengend Snippet: IRF8 depletion suppresses caspase activation and caspase activation is required for EBV lytic replication. A. IRF8 depletion suppresses caspase activation. Western blot analysis of protein extracts from Fig 1D using antibodies against caspase-3, cleaved caspase-3, caspase-8, cleaved caspase-8, caspase-7, cleaved caspase-7, caspase-9, cleaved caspass-9, caspase-2 and Bcl-2 as indicated. B. Caspase inhibition suppresses EBV lytic gene expression. Akata (EBV + ) cells were untreated or pre-treated with pan-caspase inhibitor (Z-VAD-FMK) for 1 hr and then anti-IgG was added for 48 hrs. RNA was extracted and EBV lytic gene expression was analyzed by RT-qPCR. Data are presented as means ± standard deviations of triplicate assays. ** p
Article Snippet: Membranes were then incubated in the following primary antibodies: mouse anti-ZTA (Argene, Cat # 11–007, 1:5,000), mouse anti-RTA (Argene, 1:1,000), mouse anti-BGLF4 antibody (1:1,000) [ ], anti-β-actin (Sigma, Cat # A5441, 1:5,000), anti-IRF8 (CST, Cat #5628, 1:1,000), anti-PARP (CST, Cat #9532, 1:1,000), anti-Cleaved PARP (CST, Cat #5625, 1:1,000), anti-Cleaved Caspase Substrates (CST, Cat #8698, 1:1,000), anti-Caspase-1 (CST, Cat #3866, 1:1,000), anti-Caspase-2 (CST, Cat #2224, 1:1,000), anti-Caspase-3 (Santa Cruz, Cat #sc-7148, 1:1,000), anti-Cleaved Caspase-3 (CST, Cat #9664, 1:1,000), anti-Caspase-7 (CST, Cat #12827, 1:1,000), anti-Cleaved Caspase-7 (CST, Cat #8438, 1:1,000), anti-Caspase-8 (CST, Cat #9746, 1:1,000), anti-Cleaved Caspase-8 (CST, Cat #9496, 1:1,000), anti-Caspase-9 (CST, Cat #9508, 1:1,000), anti-Bcl-2 (Bethyl, Cat #A303-675A, 1:1,000), anti-KAP1 (CST, Cat #4123, 1:1,000), anti-PAX5 (CST, Cat #8970, 1:1,000), anti-DNMT3A (Bethyl, Cat #A304-278A, 1:1,000), anti-STAT3 (CST, Cat #9139, 1:1,000), and anti-HA (CST, Cat #14031S, 1:1,000).
Techniques: Activation Assay, Western Blot, Inhibition, Expressing, Quantitative RT-PCR