Fig 11

(A and B) TMEM33-ΔTM1 and TMEM33-ΔTM2 have no effect on poly I:C/SVCV-induced IFNφ1pro/ISRE-Luc activation. EPC cells were seeded in 24-well plates and transfected the next day with 250 ng IFNφ1pro-Luc (A) or ISRE-Luc (B) and 25 ng pRL-TK, plus 250 ng TMEM33-Myc, TMEM33-ΔTM1-Myc, TMEM33-ΔTM2-Myc, TMEM33-ΔTM3-Myc, or pCMV-Myc (control vector). At 24 h post-transfection, cells were untreated (null) or transfected with poly I:C (1 μg/ml) or treated with SVCV (MOI = 1). Luciferase activities were monitored at 24 h after stimulation. The promoter activity is presented as relative light units (RLU) normalized to Renilla luciferase activity. (C and D) TMEM33-ΔTM1 and TMEM33-ΔTM2 have no effect on RLRs-induced IFNφ1pro/ISRE-Luc activation. EPC cells were seeded in 24-well plates and transfected the next day with 250 ng IFNφ1pro-Luc (C) or ISRE-Luc (D) and 250 ng MAVS-Myc, MITA-Myc, or TBK1-Myc and 25 ng pRL-TK, plus 250 ng TMEM33-Myc, TMEM33-ΔTM1-Myc, TMEM33-ΔTM2-Myc, TMEM33-ΔTM3-Myc, or pCMV-Myc (control vector). Luciferase activities were monitored at 24 h after transfection. (E-I) TMEM33-ΔTM1 and TMEM33-ΔTM2 have no effect on poly I:C/SVCV-induced the expression of ifn (E), vig1 (F), isg15-1 (G), irf7 (H), and rig-i (I). EPC cells seeded in 6-well plates overnight were transfected with the indicated plasmids (2 μg each) for 24 h. At 24 h post-transfection, cells were untreated (null) or transfected with poly I:C (1 μg/ml) or stimulated with SVCV (MOI = 1) for 24 h. 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 transcription of β-actin and 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).