Fig 3

(A and B) Enhance of virus replication by overexpression of TMEM33. EPC cells seeded in 24-well plates overnight were transfected with 0.5 μg of TMEM33-Myc 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). 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). (C-G) Overexpression of TMEM33 inhibits the expression of ifn (C), vig1 (D), isg15-1 (E), irf7 (F), and rig-i (G) induced by SVCV infection. EPC cells seeded in 6-well plates overnight were transfected with 2 μg of TMEM33-Myc or empty vector and infected with SVCV (MOI = 1) at 24 h post-transfection. At 24 h after infection, total RNAs were extracted to examine the mRNA levels of cellular ifn, vig1, isg15-1, irf7, and rig-i. 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 values (*, p < 0.05).