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Fig. 8 Overexpression of TMEM47 attenuates the antiviral function of MAVS and STING. (A–D) Overexpression of TMEM47 counteracts the antiviral effect of MAVS and STING. EPC cells seeded in 24-well plates overnight were transfected with 0.5 µg MAVS-Myc/STING-Myc plus 0.5 µg TMEM47-HA or empty vector. At 24 h post-transfection, cells were infected with SVCV (MOI = 0.001) for 48 h. Then, cells were fixed with 4% PFA and stained with 1% crystal violet (A and C). Culture supernatants from the cells infected with SVCV were collected, and the viral titer was measured according to the method of Reed and Muench (B and D). (E and F) The effect of STING and MAVS on inhibiting SVCV amplification is blocked by TMEM47. EPC cells were seeded in six-well plates overnight and transfected with indicated plasmids. At 24 h post-transfection, the cells were infected with SVCV (MOI = 1) for 24 h. After 24 h infection, the cells were harvested for IB with the indicated Abs. (G and H) The same samples were prepared similarly as described above for panels (E) and (F). Total RNAs were extracted to examine the mRNA levels of g, l, m, n, p. The relative transcriptional levels were normalized to the transcriptional level of the β-actin gene and were represented as fold induction relative to the transcriptional level in the control cells, which was set to 1. Data were expressed as mean ± SEM, n= 3. Asterisks indicate significant differences from control (*P < 0.05). All experiments were repeated for at least three times with similar results.