Journal: PLoS ONE

Article Title: Mild Endoplasmic Reticulum Stress Promotes Retinal Neovascularization via Induction of BiP/GRP78

doi: 10.1371/journal.pone.0060517

Figure Lengend Snippet: HRMEC were treated with 10 ng/ml of tunicamycin for 2 h. (A) BiP on the cell surface was precipitated after biotinylation to detect cell surface protein in untreated (control) or tunicamycin treated cells. (B) BiP and T-cadherin were immunoprecipitated with the anti-KDEL antibody to confirm the association of BiP with T-cadherin. Whole lysate without immunoprecipitation with the anti-BiP antibody was similarly evaluated. (C) After the membrane extraction, BiP and T-cadherin were immunoprecipitated with the anti-T-cadherin antibody and detected by using Western blotting with the anti-BiP antibody. Whole lysate without immunoprecipitation with the anti-VEGF receptor-2 (VEGFR-2) antibody was similarly evaluated to confirm the success of the membrane extraction. (D) Immunostaining showed the presence of BiP and T-cadherin on the surface of HRMEC. Scale bar indicates 30 µm.

Article Snippet: Immunoprecipitation products were analyzed by Western blotting using anti-BiP antibody (BD Biosciences, San Diego, CA, USA) and anti-T cadherin antibody (AnaSpec, Fremont, CA, USA).

Techniques: Immunoprecipitation, Western Blot, Immunostaining

Journal: PLoS ONE

Article Title: Mild Endoplasmic Reticulum Stress Promotes Retinal Neovascularization via Induction of BiP/GRP78

doi: 10.1371/journal.pone.0060517

Figure Lengend Snippet: (A) OIR model mice at P17, treated with 3 µg/ml of tunicamycin at P14, were perfused with FITC-dextran and the retinas were stained with anti-BiP antibody. The original images (green channel) and BiP stained images (red channel) are shown, along with the analyzed images. (B) BiP (green channel) and CD31 (red channel) staining in flat-mounted retinas of P17 mice, under normoxia or subjected to the OIR protocol, following tunicamycin injection. Arrowheads and arrows indicate normal vessels and newly formed vessels anterior to internal limiting membrane, respectively. Scale bars indicate 50 µm. The complexes of BiP and T-cadherin in the retina were immunoprecipitated with the anti-T-cadherin antibody and detected by using Western blotting with the anti-BiP antibody (C), or by the Coomassie-staining (D).

Article Snippet: Immunoprecipitation products were analyzed by Western blotting using anti-BiP antibody (BD Biosciences, San Diego, CA, USA) and anti-T cadherin antibody (AnaSpec, Fremont, CA, USA).

Techniques: Staining, Injection, Immunoprecipitation, Western Blot

Journal: STAR Protocols

Article Title: CyTOF mass cytometry analysis of human memory CD4 + T cells and memory B cells

doi: 10.1016/j.xpro.2022.101269

Figure Lengend Snippet:

Article Snippet: 161 Dy-conjugated anti-T-bet (clone 4B10) (1:50 dilution) , Fluidigm , Cat#3161014B.

Techniques: Recombinant, Saline, Staining, Software, Cytometry