Journal: Cancer Science

Article Title: Measles virus selectively blind to signaling lymphocyte activity molecule has oncolytic efficacy against nectin‐4‐expressing pancreatic cancer cells

doi: 10.1111/cas.13064

Figure Lengend Snippet: Expression of measles virus receptor molecules on the surface of pancreatic cancer cell lines. (a) Cells were incubated with anti‐nectin‐4 goat polyclonal antibody (gray histogram) or isotype control (black histogram) followed by incubation with Alexa 488‐conjugated rabbit anti‐goat antibody. (b) Nectin‐4 expression is presented as the mean fluorescence intensity (MFI) value by deducting the MFI obtained with isotype control. (c) Cells incubated with anti‐CD46 mouse mAb (red), anti‐signaling lymphocyte activity molecule (SLAM) mouse mAb (blue), or isotype control (black) followed by incubation with Alexa 488‐conjugated goat anti‐mouse antibody.

Article Snippet: The expression of nectin‐4, SLAM, and CD46 was analyzed by flow cytometry according to previously reported methods., The following antibodies were used: anti‐human SLAM mAb (clone 7D4; Biolegend, San Diego, CA, USA); anti‐human CD46 mAb, anti‐nectin‐4 goat polyclonal antibody, mouse control IgG1, and goat control IgG (all from R&D Systems, Minneapolis, MN, USA); and Alexa 488‐conjugated anti‐mouse or anti‐goat antibody (Invitrogen).

Techniques: Expressing, Virus, Incubation, Control, Fluorescence, Activity Assay

Journal: PLoS ONE

Article Title: Differential Expression of Complement Markers in Normal and AMD Transmitochondrial Cybrids

doi: 10.1371/journal.pone.0159828

Figure Lengend Snippet: The classical pathway is triggered by interaction of C1 with immune or non-immune complexes leading to conformational changes in C1q, activation of C1r and C1s, and subsequent assembly of C3 convertase. The lectin pathway is activated by binding of mannose-binding lectin (MBL) to mannose residues on the pathogen surface, which activates the MBL-associated serine proteases (MASPs), followed by formation of C3 convertase. Spontaneous hydrolysis of C3 initiates the alternative complement pathway. All three pathways of the complement cascade converge on the classical C3 convertase, which cleaves and activates component C3, forming C3a and C3b. This triggers a series of further cleavage and activation events, leading to cleavage of C5 into C5a and C5b, and eventual formation of the membrane attack complex (MAC), consisting of C5b, C6, C7, C8, and C9. MAC is the terminal cytolytic complex of the complement pathway; it causes osmotic lysis of target cells by forming transmembrane channels that disrupt the phospholipid bilayer of target cells. The complement system is tightly regulated by the following complement control proteins: CFH, CFHR1, CFHR4, CFB, CD55, CD59, CD46, CFI, and CFP.

Article Snippet: 7 , CD46 , 44 kDa , Goat Anti-CD46 Polyclonal antibody # AF2005 (RD Systems) , 1:1000 , Human , Dnk pAb to Goat IgG (HRP) ab97120 (ABCAM) , beta-actin antibody GTX 110564 (Genetex).

Techniques: Activation Assay, Binding Assay, Lysis

Journal: PLoS ONE

Article Title: Differential Expression of Complement Markers in Normal and AMD Transmitochondrial Cybrids

doi: 10.1371/journal.pone.0159828

Figure Lengend Snippet: Complement Markers’ Information.

Article Snippet: 7 , CD46 , 44 kDa , Goat Anti-CD46 Polyclonal antibody # AF2005 (RD Systems) , 1:1000 , Human , Dnk pAb to Goat IgG (HRP) ab97120 (ABCAM) , beta-actin antibody GTX 110564 (Genetex).

Techniques: Marker, Activation Assay, Lysis, Inhibition

Journal: PLoS ONE

Article Title: Differential Expression of Complement Markers in Normal and AMD Transmitochondrial Cybrids

doi: 10.1371/journal.pone.0159828

Figure Lengend Snippet: Complement Genes’ Information.

Article Snippet: 7 , CD46 , 44 kDa , Goat Anti-CD46 Polyclonal antibody # AF2005 (RD Systems) , 1:1000 , Human , Dnk pAb to Goat IgG (HRP) ab97120 (ABCAM) , beta-actin antibody GTX 110564 (Genetex).

Techniques: Variant Assay

Journal: PLoS ONE

Article Title: Differential Expression of Complement Markers in Normal and AMD Transmitochondrial Cybrids

doi: 10.1371/journal.pone.0159828

Figure Lengend Snippet: Complement Proteins’ Western Blotting Information.

Article Snippet: 7 , CD46 , 44 kDa , Goat Anti-CD46 Polyclonal antibody # AF2005 (RD Systems) , 1:1000 , Human , Dnk pAb to Goat IgG (HRP) ab97120 (ABCAM) , beta-actin antibody GTX 110564 (Genetex).

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Differential Expression of Complement Markers in Normal and AMD Transmitochondrial Cybrids

doi: 10.1371/journal.pone.0159828

Figure Lengend Snippet: Complement Gene Expression Data.

Article Snippet: 7 , CD46 , 44 kDa , Goat Anti-CD46 Polyclonal antibody # AF2005 (RD Systems) , 1:1000 , Human , Dnk pAb to Goat IgG (HRP) ab97120 (ABCAM) , beta-actin antibody GTX 110564 (Genetex).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Differential Expression of Complement Markers in Normal and AMD Transmitochondrial Cybrids

doi: 10.1371/journal.pone.0159828

Figure Lengend Snippet: Decreased protein levels of CFH, CFHL1, CD55, CD59, CFI, and CD46, and increased levels of CFP, CFB, CFHR4, and CFHR1 protein in AMD cybrids. (A, C, E, G, I, K, M, O, Q, S) Representative Western blots of CFH, CFHL1, CD55, CD59, CFI, CFP, CFB, CD46, CFHR4, and CFHR1 respectively. (B, D, F, H, J, L, N, P, R, T) Graphs showing quantitation of CFH, CFHL1, CD55, CD59, CFI, CFP, CFB, CD46, CFHR4, and CFHR1 proteins in Older-Normal and AMD cybrids. * P < 0.05, ** P < 0.01. n = 4–5. Data were analyzed using Student’s T-test.

Article Snippet: 7 , CD46 , 44 kDa , Goat Anti-CD46 Polyclonal antibody # AF2005 (RD Systems) , 1:1000 , Human , Dnk pAb to Goat IgG (HRP) ab97120 (ABCAM) , beta-actin antibody GTX 110564 (Genetex).

Techniques: Western Blot, Quantitation Assay

Journal: PLoS ONE

Article Title: Differential Expression of Complement Markers in Normal and AMD Transmitochondrial Cybrids

doi: 10.1371/journal.pone.0159828

Figure Lengend Snippet: Complement Protein Expression Data.

Article Snippet: 7 , CD46 , 44 kDa , Goat Anti-CD46 Polyclonal antibody # AF2005 (RD Systems) , 1:1000 , Human , Dnk pAb to Goat IgG (HRP) ab97120 (ABCAM) , beta-actin antibody GTX 110564 (Genetex).

Techniques: Expressing

Journal: Journal of Virology

Article Title: Neutralizing antibodies with neurotropic factor treatment maintain neurodevelopmental gene expression upon exposure to human cytomegalovirus

doi: 10.1128/jvi.00696-23

Figure Lengend Snippet: NPCs and cerebral organoids express several receptors required for HCMV entry at the plasma membrane. (A) Schematic made in BioRender depicting the HCMV viral particle noting the trimeric (gHgLgO) and pentameric (gHgLpUL128-131A) glycoprotein complexes and target cellular receptors PDGFRα and TGFβRIII (trimer) or Nrp2, THBD, and CD46 (pentamer). (B) RNA expression levels of these entry receptors as determined by analyzing previously performed bulk RNA-seq analysis in cerebral organoids (11). (C) Immunostaining conducted in passage 3 NPCs 3 days post plate down labeling receptors (PDGFRα, Nrp2, and TGFβRII) in green, membrane marker (red), Hoescht (blue), and merged to show the co-localization of membrane and receptor targets. This staining was conducted without permeabilizing the cells to strictly focus on surface expression of the receptors.

Article Snippet: The primary antibodies used were as follows: Nrp2 (R&D Systems, AF2215-SP), PDGFRα (BD Biosciences, 556001), CD46 (R&D Systems, AF2005), THBD (AbCam, ab33513), TGFβRIII (Sigma, T1940), UL123 (Shenk Lab, Clone 1B12), SOX2 (Sigma, AB5603MI), Tuj1 (GeneTex, GTX85469), and nuclear stain Hoescht (Thermo Scientific, H3570), along with a conjugated Cellbrite Orange Cytoplasmic Membrane Dye (Biotium, 30022) in .

Techniques: Clinical Proteomics, Membrane, RNA Expression, RNA Sequencing, Immunostaining, Labeling, Marker, Staining, Expressing

Journal: Heliyon

Article Title: Proteolytic shedding of CD46 from human hepatocytes indicates liver stress

doi: 10.1016/j.heliyon.2024.e40841

Figure Lengend Snippet: Establishing and validating an ELISA for soluble CD46 (sCD46) . (A) Signal:noise (S:N) values for different concentrations of CD46 capture and detection antibodies in Reagent Diluent 2 (n = 2). (B) Titration curve using recombinant human CD46 (rhCD46) as the analyte. Lower limit of quantification (LLOQ), upper limit of quantification (ULOQ) and limit of detection (LOD) are indicated (n = 3). (C) Accuracy (spike recovery) of human serum samples (n = 5). (D) Intra-assay precision, inter-assay precision and reproducibility of human serum sample at two different dilution factors. (E) Analysis of assay linearity (n = 4, Pearson correlation, indicating 90 % prediction bands). (F) Linear regression of the bias plotted against nominal titres (n = 4, Pearson correlation). (G) Robustness based on incubation timing (n = 4, one sample t -test to a mean of 1). (H) Stability of sCD46 (pg/ml) in representative cell culture supernatants over multiple freeze/thaw cycles depending on the storage temperature and the application of a protease inhibitor. (I) rhCD46 titration in ELISA and competitive FACS-based assay (n = 3). (J) Performance of commercial CD46 ELISA kits in detecting serum sCD46 compared to our in-house ELISA. Samples dilutions were based on manufacturers' recommendations.

Article Snippet: Next, 100 μl/well of samples, the corresponding 1:2 serial diluted standards of rhCD46 (2 ng/ml to 31.25 pg/ml, 10256-CD, R&D Systems) and a blank control are added and then incubated for 2 h. Then, 100 μl/well detection antibody (50 ng/ml, BAF2005, R&D Systems) is added followed by another incubation for 2 h. Next, 100 μl/well of Streptavidin-HRP (DY998, R&D Systems) is added followed by 100 μl/well of substrates (DY999, R&D Systems) for 20 and 30 min, respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Titration, Recombinant, Intra Assay, Inter Assay, Incubation, Cell Culture, Protease Inhibitor

Journal:

Article Title: Distinct Proinflammatory Host Responses to Neisseria gonorrhoeae Infection in Immortalized Human Cervical and Vaginal Epithelial Cells

doi: 10.1128/IAI.69.9.5840-5848.2001

Figure Lengend Snippet: Immunocytochemical analysis of adhesion molecule expression by cervicovaginal epithelial cells in vitro and in vivo. Positive cells appear red. (A and B) Constitutive expression of CD46 in endocervical tissue (A) and endocervical cell culture (B). (C to G) Expression of CD66 in vaginal tissue (C), ectocervical tissue (D), endocervical tissue (E), uninfected endocervical cell culture (F), and endocervical cell culture after 8 h of infection with the N. gonorrhoeae F62 piliated variant (G). (H to K) Expression of ICAM-1 in endocervical cell cultures with no infection (H), following 8 h of TNF-α stimulation (I), following 8 h of infection with N. gonorrhoeae piliated F62 (J), or following 8 h of infection with N. gonorrhoeae nonpiliated F62 (K). L. lumenal epithelial surface: B. basal epithelial layers. Magnification, ×125 (A, C to E, and H to K) and ×250 (B, F, and G).

Article Snippet: Anti-human CD46 antibody (mouse monoclonal MAS657) was purchased from Accurate Chemical and Scientific Corporation, Westbury, N.Y. Fluorescein isothiocyanate (FITC)-conjugated pan-CD66 antibody (mouse monoclonal F7112) recognizing an epitope common to four members of the CD66 family (CD66a, -b, -c, and -e) was purchased from Dako, Carpinteria, Calif.

Techniques: Expressing, In Vitro, In Vivo, Cell Culture, Infection, Variant Assay