Journal: Virus Research

Article Title: Single amino acid substitution at position 614 in SARS-CoV-2 Spike Protein alters viral assembly and infectivity

doi: 10.1016/j.virusres.2025.199624

Figure Lengend Snippet: Effect of amino acid mutation at position 614 of S protein on viral packaging. (A and B) Western blot analysis of the protein expression of pseudovirus S protein and p24 in packaging cells and virus supernatant. (C) Schematic outline of the SARS-CoV-2 VLP production process. Plasmids encoding the SARS-CoV-2 structural proteins E, M, and N were cotransfected into HEK293T cells with plasmids encoding WT S or its mutants to obtain VLPs. (D and E) Western blot analysis of the expression of the major structural proteins S and NP on the VLPs. The original image is Supplemental Figure 4 in the supplementary material.

Article Snippet: GoldBand Plus 3-color Regular Range Protein Marker(8–180 kDa) (YEASEN, #20350ES72); SARS-CoV-2 (2019-nCoV) Spike RBD Antibody, Rabbit PAb, Antigen Affinity Purified (Sinobiological, # 40,592-T62); SARS-CoV-2 (2019-nCoV) Spike S2 Antibody, Rabbit PAb, Antigen Affinity Purified (Sinobiological, #40,590-T62); GFP (4B10) Mouse mAb (CST, #2955); Flag (DYKDDDDK) tag Polyclonal Antibody (Proteintech, #20,543–1-AP); Human Immunodeficiency Virus type 1 (HIV-1) Gag-p24 Antibody, Rabbit PAb, Antigen Affinity Purified (Sinobiological, #11,695-RP02); Myc-Tag (9B11) Mouse mAb (CST, #2276); GAPDH Mouse Monoclonal Antibody (Origene, #OTI2D9); β-actin Polyclonal Antibody (Proteintech, #20,536–1-AP).

Techniques: Mutagenesis, Western Blot, Expressing, Virus

Journal: The Journal of Biological Chemistry

Article Title: Externalized Glycolytic Enzymes Are Novel, Conserved, and Early Biomarkers of Apoptosis *

doi: 10.1074/jbc.M111.314971

Figure Lengend Snippet: Underrepresented apoptotic membrane-associated proteins Proteins identified by iTRAQ analysis and found to be underrepresented among apoptotic membrane vesicles by at least 20% at a confidence level of >95% are listed.

Article Snippet: Human transformed (Jurkat) T lymphocytes that had been induced to undergo apoptosis by treatment with actinomycin D or that had been left untreated were analyzed cytofluorometrically following staining with FITC-conjugated mouse anti-human annexin II monoclonal antibody (BD Biosciences) ( A ) or mouse anti-calreticulin monoclonal and FITC-conjugated goat anti-rabbit IgG secondary antibodies (Enzo Life Sciences) ( B ).

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: Externalized Glycolytic Enzymes Are Novel, Conserved, and Early Biomarkers of Apoptosis *

doi: 10.1074/jbc.M111.314971

Figure Lengend Snippet: Characterization of appearance of exposed glycolytic enzyme molecules. Murine splenocytes that were induced to undergo apoptosis with staurosporine were analyzed cytofluorometrically following staining with PE-conjugated annexin V, 7-AAD, and FITC-conjugated goat anti-EnoA peptide IgG polyclonal antibody (A), rabbit anti-GAPDH IgG polyclonal antibody and FITC-conjugated goat anti-rabbit IgG secondary antibody (B), and rabbit anti-TPI IgG polyclonal antibody and FITC-conjugated goat anti-rabbit IgG secondary antibody (C). Cells that met the criteria of staining positively with annexin V and negatively with 7-AAD (Annexin V+ 7-AAD−; the R1 region indicated in red in the upper dot plots) were gated electronically, and the fluorescein signal of those cells was analyzed (shown as solid green histograms in the lower panels). The fluorescein signal of annexin V+/7-AAD− cells stained with secondary antibody alone also is presented (gray dotted lines).

Article Snippet: Human transformed (Jurkat) T lymphocytes that had been induced to undergo apoptosis by treatment with actinomycin D or that had been left untreated were analyzed cytofluorometrically following staining with FITC-conjugated mouse anti-human annexin II monoclonal antibody (BD Biosciences) ( A ) or mouse anti-calreticulin monoclonal and FITC-conjugated goat anti-rabbit IgG secondary antibodies (Enzo Life Sciences) ( B ).

Techniques: Staining

Journal: The Journal of Biological Chemistry

Article Title: Externalized Glycolytic Enzymes Are Novel, Conserved, and Early Biomarkers of Apoptosis *

doi: 10.1074/jbc.M111.314971

Figure Lengend Snippet: Cytofluorometric analysis of plasminogen binding to apoptotic cells. A, murine splenocytes that had undergone apoptosis spontaneously in culture (12 h) and freshly isolated, viable splenocytes were analyzed cytofluorometrically as described in the legend of Fig. 6 following staining with FITC-conjugated plasminogen. B, murine splenocytes that were induced to undergo apoptosis with staurosporine were analyzed cytofluorometrically following staining with PE-conjugated annexin V, 7-AAD, and FITC-conjugated plasminogen and analyzed as described in the legend of Fig. 8.

Article Snippet: Human transformed (Jurkat) T lymphocytes that had been induced to undergo apoptosis by treatment with actinomycin D or that had been left untreated were analyzed cytofluorometrically following staining with FITC-conjugated mouse anti-human annexin II monoclonal antibody (BD Biosciences) ( A ) or mouse anti-calreticulin monoclonal and FITC-conjugated goat anti-rabbit IgG secondary antibodies (Enzo Life Sciences) ( B ).

Techniques: Binding Assay, Isolation, Staining

Journal: The Journal of Biological Chemistry

Article Title: Externalized Glycolytic Enzymes Are Novel, Conserved, and Early Biomarkers of Apoptosis *

doi: 10.1074/jbc.M111.314971

Figure Lengend Snippet: Cytofluorometric analysis of externalization of other molecules. Human transformed (Jurkat) T lymphocytes that had been induced to undergo apoptosis by treatment with actinomycin D or that had been left untreated were analyzed cytofluorometrically following staining with FITC-conjugated mouse anti-human annexin II monoclonal antibody (BD Biosciences) (A) or mouse anti-calreticulin monoclonal and FITC-conjugated goat anti-rabbit IgG secondary antibodies (Enzo Life Sciences) (B). Apoptotic (solid green histograms) and viable (dashed lines) cells were identified by scatter properties and gated electronically.

Article Snippet: Human transformed (Jurkat) T lymphocytes that had been induced to undergo apoptosis by treatment with actinomycin D or that had been left untreated were analyzed cytofluorometrically following staining with FITC-conjugated mouse anti-human annexin II monoclonal antibody (BD Biosciences) ( A ) or mouse anti-calreticulin monoclonal and FITC-conjugated goat anti-rabbit IgG secondary antibodies (Enzo Life Sciences) ( B ).

Techniques: Transformation Assay, Staining