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

Journal: Nutrients

Article Title: Vitamin A Status Modulates Epithelial Mesenchymal Transition in the Lung: The Role of Furin.

doi: 10.3390/nu16081177

Figure Lengend Snippet: Figure 3. Effect of RA addition in vitro in human lung cell lines. (A) Western blot analysis showed the expression of different EMT markers and furin in two cell lines (A549 and BEAs) after incubation with 5 µM RA at 24 h and 48 h. Hsc70 was used as a loading control. A representative image is shown (n = 3 independent cultures). (B) Localization of E-cadherin (green), detected by immunofluorescence staining in A549 and BEA cells after 5 µM RA treatment for 24 h. Zoom has been made around the dotted lines and magnified images are shown on the right side. (C) Representative IF analysis of furin (green) in lung cell lines (A549 and BEAs) after 24 h of RA incubation (5 µM). In both (B,C), nuclei were stained with DAPI and F-actin with phalloidin (cyan blue). Scale bars 20 µM and 60 µM for zoom images in (B).

Article Snippet: Primary antibodies against Furin (70393), Vimentin (5741), N-cadherin (14215), and β-Actin (4970) were purchased from Cell Signalling Technology (Danvers, MA, USA).

Techniques: In Vitro, Western Blot, Expressing, Incubation, Control, Immunofluorescence, Staining

Journal: Nutrients

Article Title: Vitamin A Status Modulates Epithelial Mesenchymal Transition in the Lung: The Role of Furin.

doi: 10.3390/nu16081177

Figure Lengend Snippet: Figure 5. RARα ChIP assay on furin promoter. (A) Potential binding sites for RAR and/or RXR nuclear receptors in the furine promoter region. CTF1, CTF2, and Enhancer (ENH) indicate 3 regions where most transcriptional factor binding sites that modulate gene transcription are concentrated. (B) ChIP data analysis of RARα binding to furin and MMP-9 genes in control rats (RA−) and control rats injected intraperitoneally with retinoic acid (RA+) (left panel, CONTROL group). The same analysis was carried out in vitamin A-deficient rats (RA−) and vitamin A-deficient rats injected intraperitoneally with retinoic acid (RA+) (Right panel, VAD). Chromatin was immunoprecipitated with an anti-RARα antibody (black bars) or a nonrelated antibody anti-IgG as a negative control (grey bars). Data are the result of three independent experiments and bars represent mean ± SD.

Article Snippet: Primary antibodies against Furin (70393), Vimentin (5741), N-cadherin (14215), and β-Actin (4970) were purchased from Cell Signalling Technology (Danvers, MA, USA).

Techniques: Binding Assay, Control, Injection, Immunoprecipitation, Negative Control