Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Nervous necrosis virus capsid protein functions as a SUMO E3 ligase to activate MAVS-dependent NF-κB signaling

doi: 10.1007/s00018-026-06155-8

Figure Lengend Snippet: CP does not interfere with MAVS localization to mitochondria. HEK 293T cells ( A ) or FHM cells ( B )were co-transfected with pCMV-Flag-MAVS and pCMV-Myc-CP plasmids, and analyzed by immunofluorescence (IF) assays. CP was detected using anti-Myc antibody (magenta) and MAVS with anti-Flag antibody (green). Mitochondria were labeled with MitoTracker Red (red), and nuclei were stained with DAPI (blue). Fluorescence intensity profiles were quantified using ImageJ across representative confocal fields. Scale bar = 10 μm

Article Snippet: Primary antibodies included rabbit polyclonal anti-UBC9 ( T55571 ), mouse monoclonal anti-Myc ( M20002 ), anti-HA ( M20003 ), anti-Flag (M20008), anti-β-actin ( M20011 ), and anti-His ( M30111 ), all obtained from Abmart (Shanghai, China).

Techniques: Transfection, Immunofluorescence, Labeling, Staining, Fluorescence

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Nervous necrosis virus capsid protein functions as a SUMO E3 ligase to activate MAVS-dependent NF-κB signaling

doi: 10.1007/s00018-026-06155-8

Figure Lengend Snippet: CP interacts with SUMO isoforms and the E2-conjugating enzyme UBC9. ( A ) Subcellular co-localization of CP with SUMO1, SUMO2, and SUMO3. HEK 293T cells were co-transfected with pCMV-Flag-CP and pCMV-HA-SUMO1/2/3 plasmids for IF assays, using anti-HA (green), anti-Flag (red), and DAPI (blue) for nuclear staining. Co-localization signals were visualized by confocal microscopy and quantified using ImageJ. Scale bar = 10 μm. (B-C) Physical interaction between CP and SUMO isoforms. HEK 293T cells were co-transfected with pCMV-Flag-CP or pCMV-Myc-CP and pCMV-HA-SUMO1/2/3 plasmids. Co-IP was performed using anti-Flag magnetic beads ( B ) or anti-HA magnetic beads ( C ), followed by immunoblotting. ( D ) Subcellular co-localization of CP with UBC9. HEK 293T cells were co-transfected with pCMV-Flag-CP and pCMV-Myc-UBC9 . IF assays were conducted using anti-Myc (green), anti-Flag (red), and DAPI (blue). Confocal imaging and ImageJ quantification were used to assess co-localization. Scale bar = 10 μm. ( E ) Co-IP analysis of the CP–UBC9 interaction. HEK 293T cells were co-transfected with pCMV-Flag-CP and pCMV-Myc-UBC9 . Lysates were immunoprecipitated with anti-Flag magnetic beads and analyzed by immunoblotting. ( F ) Pull-down analysis of CP and UBC9 protein interaction. HEK 293T cells were transfected with pCMV-Myc-UBC9 plasmids and pull-down assays were performed with pET32a-His-CP protein lysates. Lysates were incubated with anti-His magnetic beads for immunoprecipitation. Associated proteins were detected by immunoblotting

Article Snippet: Primary antibodies included rabbit polyclonal anti-UBC9 ( T55571 ), mouse monoclonal anti-Myc ( M20002 ), anti-HA ( M20003 ), anti-Flag (M20008), anti-β-actin ( M20011 ), and anti-His ( M30111 ), all obtained from Abmart (Shanghai, China).

Techniques: Transfection, Staining, Confocal Microscopy, Co-Immunoprecipitation Assay, Magnetic Beads, Western Blot, Imaging, Immunoprecipitation, Incubation