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Collection : Elife (E-Journal - Via Crossref)

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Figure 2—figure supplement 1. Active RV replication induces proteasomal degradation of MAVS. ; (A) MA104 cells were infected with RRV (MOI = 1) and harvested at indicated time points post infection for RT-qPCR analysis. MAVS mRNA levels were measured and normalized to that of GAPDH. (B) African green monkey COS-7 cells were infected with RRV (MOI = 3) and harvested at indicated time points for western blot (top panel). MA104 cells were infected with different RV strains (MOI = 3) and harvested at 8 hpi for western blot (middle panel). HT-29 cells were infected with Wa at MOI = 1 or 10 for indicated time points and harvested for western blot. (RRV, SA11: simian RVs; UK: bovine RV; OSU, SB1A: porcine RVs; Wa, DS1: human RVs; ETD: murine RV). (C) MA104 cells were infected with RRV (MOI = 3), with RRV mixed with neutralizing antibodies (nAb) or with psoralen UV-inactivated RRV (UV) for 6 or 12 hpi. The lysates were harvested and examined by western blot. (D) Supernatants were harvested from mock-infected MA104 cells (sup(M)) or RRV-infected MA104 cells (sup(R)) and used to re-infect fresh MA104 cells. Lysates were harvested at 6 or 12 hpi and examined by western blot. (E) MA104 cells were infected with RRV (MOI = 3) for 12 hpi in the presence of different chemical inhibitors. (ITZ: itraconazole; Salu: salubrinal; EerI: Eeyarestatin I; Spi: spiroepoxide; GW: GW4869; STS: staurosporine; Z-VAD: Z-VAD-FMK). (F) MA104 cells were infected with RRV (MOI = 3) and treated with MG132 at indicated time point post infection (top panel). MA104 cells were infected with RRV (MOI = 3) for 12 hpi in the presence of different chemical inhibitors. (MG: MG132; lact: lactacystin; chlo: chloroquine; conA: concanamycin A; Z-VAD: Z-VAD-FMK; 3-MA: 3-Methyladenine; rap: rapamycin). For all figures, experiments were repeated at least three times with similar results. Data are represented as mean ± SEM. Statistical significance is determined by Student’s t test (p≤0.05; p≤0.01; p≤0.001).

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