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. 2018 Jun 29;92(14):e00321-18. doi: 10.1128/JVI.00321-18

FIG 7.

FIG 7

TRIM21 promotes the activation of IRF3 and NF-κB signaling and suppresses viral infection. (A) Immunoblot analysis of the indicated proteins in HLCZ01-shcon and HLCZ01-shTRIM21 cells infected with VSV for the indicated times. (B and C) HLCZ01 cells were transfected with p3X Flag-CMV-TRIM21 or pSilencer-TRIM21 for 36 h and then infected with NDV (B) or SeV (C) for 16 h. The dimer and monomer of IRF3 were detected by native PAGE. Phosphorylated IRF3 was examined by immunoblotting. (D) Huh7.5.1(MAVS-C508R) cells were transfected with pSilencer-vector or pSilencer-TRIM21 for 24 h and then infected with HCV (MOI = 2) for 48 h. Phosphorylated IRF3, phosphorylated P65, and HCV core protein were detected by immunoblotting. (E to G) HLCZ01 cells were transfected with pSilencer-vector or pSilencer-TRIM21 for 24 h and then infected with VSV (E), NDV (F), or SeV (G) for the indicated times. RNA of VSV, NDV, and SeV was analyzed by real-time PCR and normalized with GAPDH. (H) Huh7.5.1(MAVS-C508R) cells were transfected with pSilencer-vector or pSilencer-TRIM21 for 24 h and then infected with HCV (MOI = 2) for the indicated times. HCV RNA was analyzed by real-time PCR. (I) Huh7.5.1(MAVS-C508R) cells were transfected with pSilencer-vector or pSilencer-TRIM21 for 24 h and then infected with HCV (MOI = 2) for the indicated times. Extracellular infectious virus particles were detected by FFU assay. The results are presented as means ± standard deviations. *, P < 0.05; **, P < 0.01; ***, P < 0.001 versus control.