Figure 1

Identification of the interaction between the NDV M protein and chBRD2 protein. A Fluorescence co-localization of the NDV M protein and chBRD2 protein in plasmid-co-transfected cells. The plasmids pEGFP-M and pDsRed-chBRD2 or pCMV-HA-M and pCMV-Myc-chBRD2 were co-transfected into DF-1 cells. Twenty-four hours after transfection, direct fluorescence methods and indirect immunofluorescence assays were used to observe the fluorescence of EGFP-M and DsRed-chBRD2 or HA-M and Myc-chBRD2. DAPI was used to detect nuclei. The original magnification was 1 × 200. B Identification of the interaction between the M protein and chBRD2 protein by co-IP assay. The indicated plasmids were co-transfected into DF-1 cells, and reciprocal co-IP assays were performed to identify the interaction between HA-M and Myc-chBRD2 in DF-1 cells at 36 hpt. C A Coomassie stained gel showing the bacterial expression purified proteins. The His-tagged chBRD2 (His-chBRD2), His tag, GST-tagged M (GST-M), and GST tag were expressed in E. coli BL21 (DE3), and the soluble His-chBRD2 and His tag or GST-M and GST tag were purified on Ni–NTA His*Bind Resin or Glutathione-Sepharose 4B beads, respectively. The bacterial expression purified proteins were detected by SDS-PAGE along with Coomassie blue staining. Identification of the interaction between the M protein and chBRD2 protein by His pull-down assay (D) and GST pull-down assay (E). Upper panel, the purified His-chBRD2 or GST-M (Input) and the pull-downed His-chBRD2 or GST-M were detected by western blot using a mouse anti-His or anti-GST antibody. Lower panel, the bait proteins (GST and GST-M or His and His-chBRD2) in the pull-down fractions were detected by western blot using a mouse anti-GST or anti-His antibody.