primary antibodies against furin  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Cryo-electron tomography reveals coupled flavivirus replication, budding and maturation

doi: 10.1101/2024.10.13.618056

Figure Lengend Snippet: (A) Slice through tomogram of LGTV-infected cell showing an immature virion budding across from a RO (orange arrow), and mature virions (red arrows) in adjacent membranes. (B) Segmentation of the tomogram in (A), with color labels defined for each structure. (C) Slice through tomogram of LGTV-infected showing mature (red arrows) and immature (orange arrows) LGTV virions observed near the viral RO. (D-E) Subtomogram averages of immature (D) and mature (E) LGTV from cellular tomograms. (F) The distance from immature (n=37) and mature (n=24) virions to the closest RO, from five tomograms. Statistical significance by unpaired two-tailed Student’s t test: **** p < 0.005. (G) Representative immunofluorescence micrograph of dsRNA, furin and Golgi marker GM130 in LGTV-infected cells at 24 h p.i. Rightmost panel: merge including DAPI-staining of nuclei (blue). Scale bars 100 nm (A,C), 5 µm (G).

Article Snippet: The cells were then stained with primary antibodies against furin (goat polyclonal, dilution 1:50, Invitrogen), GM130 (mouse monoclonal clone 35/GM130, dilution 1:300, BD Transduction) or dsRNA (mouse monoclonal clone J2, dilution 1:1000, Scicons, Nordic MUbio; conjugated to allophycocyanin, Abcam) and secondary fluorescent antibodies (donkey anti-goat Alexa Fluor 488, dilution 1:1000, Invitrogen; goat anti-mouse Alexa Fluor 568, dilution 1:1000, Invitrogen) and DAPI diluted in blocking buffer for 1 hour each.

Techniques: Infection, Two Tailed Test, Immunofluorescence, Marker, Staining

Journal: bioRxiv

Article Title: Cryo-electron tomography reveals coupled flavivirus replication, budding and maturation

doi: 10.1101/2024.10.13.618056

Figure Lengend Snippet: Immunofluorescence microscopy of uninfected cells showing furin (A) and Golgi marker GM130 (B) and their colocalization in the absence of viral infection (D). Additional channels contain dsRNA staining (C) and DAPI staining of cell nuclei (D). Scale bars, 10 µm.

Article Snippet: The cells were then stained with primary antibodies against furin (goat polyclonal, dilution 1:50, Invitrogen), GM130 (mouse monoclonal clone 35/GM130, dilution 1:300, BD Transduction) or dsRNA (mouse monoclonal clone J2, dilution 1:1000, Scicons, Nordic MUbio; conjugated to allophycocyanin, Abcam) and secondary fluorescent antibodies (donkey anti-goat Alexa Fluor 488, dilution 1:1000, Invitrogen; goat anti-mouse Alexa Fluor 568, dilution 1:1000, Invitrogen) and DAPI diluted in blocking buffer for 1 hour each.

Techniques: Immunofluorescence, Microscopy, Marker, Infection, Staining