NS1 inhibits the ubiquitination of full-length RIG-I but not that of RIG-I 2CARD in murine cells.

<p>(<b>A</b>) Human HEK293T (<b>left</b>) or mouse Hepa 1.6 (<b>right</b>) cells were transfected with human GST-RIG-I 2CARD (GST-hRIG-I 2CARD) (<b>left</b>) or mouse GST-RIG-I 2CARD (GST-mRIG-I 2CARD) (<b>right</b>) and increasing amounts of NS1-PR8 (<b>left and right</b>). At 40 h posttransfection, WCLs were subjected to GST-pulldown (GST-PD), followed by IB with anti-GST antibody. Arrow heads indicate ubiquitinated bands. Arrow indicates non-ubiquitinated 2CARD. (<b>B</b>) Human HEK293T (<b>left</b>) or mouse Hepa1.6 (<b>right</b>) cells were transfected with Flag-tagged human RIG-I (Flag-hRIG-I) or Myc-tagged mouse RIG-I (Myc-mRIG-I) respectively, together with empty vector or HA-ubiquitin and increasing amounts of NS1-PR8. At 24 h posttransfection, cells were infected with SeV (10 HA units/ml) for 10 h. WCLs were subjected to IP with anti-Flag (<b>left</b>) or anti-Myc (<b>right</b>), followed by IB with the indicated antibodies.