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Interfering SeV replication with shRNA affects innate immune response.

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posted on 03.06.2011, 02:30 by Junqiang Ye, Tom Maniatis

A. Targeting SeV by shRNA modulates the expression of critical factors in innate immunity. MEFs were treated with shRNA specifically targeting the SeV PVC gene or a scramble sequence shRNA as control. Cells were then infected with SeV. Total protein lysates were prepared after indicated time and separated on SDS-PAGE for western blot analyses with anti-STAT1, MAVS, IRF3, SeV C, V, P and NP and β-Actin antibodies. B. Targeting SeV by shRNA has minimal effects on IFNβ expression kinetics. Experiments were conducted similarly as in A, but cells were harvested after indicated time for RNA extraction instead. The expression of IFNβ, Cxcl10, SeV NP and β-Actin was analyzed by RT-PCR.