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Image Search Results
Journal: Viruses
Article Title: PRRSV Non-Structural Proteins Orchestrate Porcine E3 Ubiquitin Ligase RNF122 to Promote PRRSV Proliferation
doi: 10.3390/v14020424
Figure Lengend Snippet: Interaction between porcine RNF122 and MDA5 molecules. ( a ) HEK293T cells were co-transfected with Flag-RNF122, 24 h after transfection, Flag Beads were used for protein enrichment, and further detected by Western blotting. Then the gel was stained with silver using a ProteoSilver Silver Stain Kit (Solarbio Life Sciences). ( b ) HEK293T cells were co-transfected with HA-RNF122 and Flag-RIG-I, Flag-MDA5 or Flag-CMV. Flag Beads were used for CO-24 h after transfection IP, and further detected by Western blotting with an anti-Flag antibody and an anti-HA antibody, respectively. ( c ) HeLa cells were placed on coverslips in 12-well plates and co-transfected with the HA-RNF122 and Flag-RIG-I, Flag-MDA5 or Flag-CMV, respectively. Fixed double-stained with a mouse anti-HA mAb and a rabbit anti-Flag antibody, and followed by FITC-conjugated anti-mouse IgG (green) and PE-conjugated anti-rabbit IgG (red). Nuclei were stained with DAPI (blue). Fluorescence confocal microscopy (UltraView Vox, PerkinElmer, Waltham, MA, USA) was used to detect the co-location. ( d ) HEK293T cells were co-transfected with HA-RNF122 and Flag-MDA5, Flag-MDA5 (N), Flag-MDA5 (M), Flag-MDA5 (C)or Flag-CMV. Flag Beads were used for CO-IP 24 h after transfection, and further detected by Western blotting with an anti-Flag antibody and an anti-HA antibody, respectively. ( e ) HEK293T cells were co-transfected with Flag-MDA5 and Myc-RNF122, Myc-RNF122 (TM), Myc-RNF122 (RING) or Myc-CMV. Flag Beads were used for CO-IP 24 h after transfection and further detected by Western blotting with an anti-Flag antibody and an anti-Myc antibody, respectively.
Article Snippet: Then the gel was stained with silver using a
Techniques: Transfection, Protein Enrichment, Western Blot, Staining, Silver Staining, Fluorescence, Confocal Microscopy, Co-Immunoprecipitation Assay