Additional file 1: Fig. S1. of G45R mutation in the nonstructural protein 1 of A/Puerto Rico/8/1934 (H1N1) enhances viral replication independent of dsRNA-binding activity and type I interferon biology

Three replicates of dsRNA-RBD pulled down assay. 0.6 μM of biotinylated-dsRNA and 0.4 mg NS1-RBDs were used in the pulled down assay. The assay was performed using similar condition in three separated reactions. The pulled-down NS1-RBD protein was electrophoresed through 15 % acrylamide gel and blotted onto a nitrocellulose membrane. Western blot was performed by using anti-NS1 rabbit serum and goat-anti rabbit IgG-HRP as primary and secondary antibodies, respectively. The protein was detected by chemiluminescent method. The G45R bound to dsRNA with lower capacity than the wild type (WT). The R38AK41A (AA) served as a negative control failed to bind with dsRNA. (TIFF 305 kb)