Table 3.

Quantitative RT-PCR Analysis of IRF5, IRF7, and IRF5 + IRF7 Overexpression in MDAH041 Cells

(A) IRF5, IRF7, and IRF5 + IRF7 Transfected vs Vector Control
041IM pCMV-IRF5041IM pCMV-IRF7041IM pCMV-IRF5 + IRF7
TLR31.22−1.88−1.84
IFNα4.352.00−1.46
IFNβ4.992.41−1.13
IRF53.101.312.19
IRF76.23657.11572.05
STAT1−2.14−1.551.39
OAS15.941.044.44
(B) Time Course of PolyI:C Treated vs Untreated

1 h
2 h
4 h
6 h
8 h
24 h
IRF5
    041IM pcDNA3.11.29−1.551.821.592.991.83
    041IM pCMV-IRF53.58−1.021.241.48−1.69−3.10
    041IM pCMV-IRF7−1.29−1.04−1.75−2.04−1.20−1.06
    041IM pCMV-IRF5 + IRF71.254.17−1.353.81−1.37−1.42
IRF7
    041IM pcDNA3.1−4.691.012.205.101.851.88
    041IM pCMV-IRF52.50−1.021.12−5.24−2.19−2.69
    041IM pCMV-IRF72.032.461.842.10−1.181.14
    041IM pCMV-IRF5 + IRF71.411.461.691.07−1.12−1.21
TLR3
    041IM pcDNA3.1−6.73−4.501.041.473.3439.40
    041IM pCMV-IRF53.39−1.401.641.711.4917.39
    041IM pCMV-IRF7−1.261.28−2.64−1.610.676.63
    041IM pCMV-IRF5 + IRF72.012.971.445.3510.2778.79
IFNα
    041IM pcDNA3.1−3.34−1.852.462.072.461.52
    041IM pCMV-IRF53.25−3.141.341.32−1.34−2.99
    041IM pCMV-IRF7−1.421.01−2.51−1.77−1.101.20
    041IM pCMV-IRF5 + IRF76.028.821.3910.565.821.91
OAS1
    041IM pcDNA3.1−2.751.68−5.3534.30187.402,019.80
    041IM pCMV-IRF52.004.005.0619.2929.24685.02
    041IM pCMV-IRF72.042.396.878.1766.72661.68
    041IM pCMV-IRF5 + IRF71.142.1412.2132.9095.67694.58
  • NOTE: The immortal MDAH041 cells were transfected with pCMV-IRF5 and/or pCMV-IRF7 and treated with 100 μg/mL polyI:C for different periods of time (0, 1, 2, 4, 6, 8, and 24 h). Expression levels of IRF5, IRF7, and IRF-inducible genes were examined by quantitative RT-PCR. 041IM, MDAH041 immortal cells. (A) Transcription levels of IRF5, IRF7, and IRF-inducible genes in IRF5-, IRF7-, and IRF5 + IRF7–transfected immortal MDAH041 cells relative to pcDNA3.1 vector control immortal MDAH041 cells. (B) Time course of transcription levels of IRF5, IRF7, and IRF-inducible genes in stably transfected cells after polyI:C treatment compared with untreated cells.