Journal: Virology Journal

Article Title: SARS-CoV-2 viral protein Nsp2 stimulates translation under normal and hypoxic conditions

doi: 10.1186/s12985-023-02021-2

Figure Lengend Snippet:

Article Snippet: Goat polyclonal anti-VEGF-C , R & D systems , Cat# AF752-SP.

Techniques: Modification, Western Blot, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Luciferase, Software, Recombinant, Imaging, Microscopy

Journal: JCI Insight

Article Title: ADAMTS2 and ADAMTS14 can substitute for ADAMTS3 in adults for pro-VEGFC activation and lymphatic homeostasis

doi: 10.1172/jci.insight.151509

Figure Lengend Snippet: ( A ) Conditioned medium from HEK293 cells expressing full-length pro-VEGFC was incubated with buffer (as negative control, lane1), ADAMTS3 (as positive control, lane 2), ADAMTS2, or ADAMTS14, in the presence or absence of EDTA used as inhibitor. The electrophoretic pattern of VEGFC was analyzed by Western blotting in reducing conditions. In absence of active enzymes (lane 1, 3, 5, and 7), VEGFC can be detected as a 58 kDa form (full-length pro-VEGFC without signal peptide) and a 31 kDa form generated by C-terminal processing by furin. In the presence of active ADAMTS3 (lane 2), ADAMTS2 (lane 4), and ADAMTS14 (lane 6), the 58 kDa form was totally converted into a 45 kDa polypeptide, whereas the 31 kDa form was processed into the fully mature 21 kDa VEGFC, which is in line with N-terminal processing of VEGFC proteins. ( B ) Schematic illustration of the different VEGFC forms, with their molecular weights provided (SP, signal peptide; NT, N-terminal propeptide; VHD, VEGF homology domain; CT, C-terminal propeptide).

Article Snippet: Western blotting analyses were performed using a polyclonal goat anti-human VEGFC primary antibody (1:250, R&D Systems, AF752).

Techniques: Expressing, Incubation, Negative Control, Positive Control, Western Blot, Generated

Journal: JCI Insight

Article Title: ADAMTS2 and ADAMTS14 can substitute for ADAMTS3 in adults for pro-VEGFC activation and lymphatic homeostasis

doi: 10.1172/jci.insight.151509

Figure Lengend Snippet: Conditioned medium from HEK293 cells expressing full-length pro-VEGFC was first incubated for 18 hours with buffer alone (lane 1, negative control), ADAMTS3 (as positive control), ADAMTS2, or ADAMTS14, in the presence or absence of EDTA used as inhibitor. These different pretreated media were then added (20 μL or 100 μL) into 1 mL of serum-free EBM-2 on LEC cultures. ( A ) After 5 minutes, cells were lysed, and phosphorylated VEGFR3 (pVEGFR3) was visualized by Western blotting. ( B ) After stripping of the antibodies, the same membrane was then used to visualize total VEGFR3. Treatment of the pro-VEGFC–rich conditioned medium with active ADAMTS3, ADAMTS2, and ADAMTS14 induced the phosphorylation of the 3 bands corresponding to VEGFR3 (arrows) in a dose-dependent manner, while the total amount of VEGFR3 was not affected, demonstrating that processing of pro-VEGFC by ADAMTS2, ADAMTS3, or ADAMTS14 leads to the activation of pro-VEGFC in a similar manner.

Article Snippet: Western blotting analyses were performed using a polyclonal goat anti-human VEGFC primary antibody (1:250, R&D Systems, AF752).

Techniques: Expressing, Incubation, Negative Control, Positive Control, Western Blot, Stripping Membranes, Membrane, Phospho-proteomics, Activation Assay