vvl fitc  (Vector Laboratories)

Journal: bioRxiv

Article Title: Avian and Human Influenza Viruses Exhibit Distinct Glycoconjugate Receptor Specificities in Human Lung Cells

doi: 10.1101/2022.06.29.498208

Figure Lengend Snippet: (A) Binding specificities of different lectins used in this study. GNL - Galanthus Nivalis Lectin, SNA - Sambucus Nigra Lectin, VVL - Vicia Villosa Lectin, and CTB - Cholera Toxin B subunit. (B) Western blot analysis of MGAT1 and C1GALT1 expression in KO cells. β-actin levels are shown as loading controls. (C) Comparison of the binding properties of various lectins in WT and KO cells. Representative flow cytometry plots showing differences in lectin binding. Data are representative of at least two independent experiments.

Article Snippet: Fluorescein labeled Galanthus Nivalis Lectin (GNL-FITC, #FL-1241 1:250), Cy3 labeled Sambucus Nigra Lectin (SNA-Cy3, #CL-1303, 1:500), and fluorescein labeled Vicia Villosa Lectin (VVL-FITC, #FL-1231, 1:500) were purchased from Vector Laboratories.

Techniques: Binding Assay, Western Blot, Expressing, Flow Cytometry

Journal: Pathogens

Article Title: The Oesophageal Squamous Cell Carcinoma Cell Line COLO-680N Fails to Support Sustained Cryptosporidium parvum Proliferation

doi: 10.3390/pathogens11010049

Figure Lengend Snippet: Exemplary illustration of Cryptosporidium parvum in vitro cell cultures at 24 hpi ( A ) Vicia villosa (VV) lectin-based detection of C. parvum (green), illustration of cell membranes via anti-β-catenin-mediated staining (red) and of cell nuclei via DAPI staining (blue). Scale bar 20 µm. ( B ) Infection rates of C. parvum -infected HCT-8 and COLO-680N cells after application of three different infection protocols. Infection rates are expressed as mean ± SD of six replicates per condition. For statistical analysis, a one-way analysis of variance (ANOVA) with Tukey’s test was performed using GraphPad ® Prism 8 software (San Diego, CA, USA), with a significance level of 5%. The significance levels are as follows: *** = p ≤ 0.001, ** = p ≤ 0.01 and presented within the table in ( B ).

Article Snippet: For detection of intracellular parasites and posterior calculation of infection rates, staining with the fluorescein labelled Vicia villosa lectin (VVL, FL-1231-2, 1:2000, VECTOR laboratories, 45 min, room temperature (RT), dark chamber) was used.

Techniques: In Vitro, Staining, Infection, Software

Journal: Pathogens

Article Title: The Oesophageal Squamous Cell Carcinoma Cell Line COLO-680N Fails to Support Sustained Cryptosporidium parvum Proliferation

doi: 10.3390/pathogens11010049

Figure Lengend Snippet: Replication of C. parvum in HCT-8- and COLO-680N cells using infection protocol III. Parasite intracellular replication was quantified by both qPCR- ( A ) and Vicia villosa (VV) lectin (VVL)-based immunofluorescence analyses ( B ). ( C ) Exemplary illustration of both in vitro cell culture systems: fluorescence-based detection of C. parvum via VVL (green) and cell nuclei via DAPI (blue). To assess parasite development, a two-tailed t -test was performed, comparing the infection rate per day, measured in HCT-8- and COLO-680N cells. The significance values were as follows: *** = p ≤ 0.001, ** = p ≤ 0.01. Scale bar 20 µm.

Article Snippet: For detection of intracellular parasites and posterior calculation of infection rates, staining with the fluorescein labelled Vicia villosa lectin (VVL, FL-1231-2, 1:2000, VECTOR laboratories, 45 min, room temperature (RT), dark chamber) was used.

Techniques: Infection, Immunofluorescence, In Vitro, Cell Culture, Fluorescence, Two Tailed Test

Journal: eLife

Article Title: The enteric pathogen Cryptosporidium parvum exports proteins into the cytosol of the infected host cell

doi: 10.7554/eLife.70451

Figure Lengend Snippet:

Article Snippet: Other , Fluorescin Vicia villosa lectin stain , Vector Labs , Cat# FL-1231-2 , IF (1:1000).

Techniques: Modification, Transgenic Assay, Expressing, Transfection, Construct, Isolation, Produced, In Vivo, Recombinant, Plasmid Preparation, Control, Sequencing, Luciferase, Software, Staining, Flow Cytometry

Journal: eLife

Article Title: The enteric pathogen Cryptosporidium parvum exports proteins into the cytosol of the infected host cell

doi: 10.7554/eLife.70451

Figure Lengend Snippet: ( A ) Schematic overview of the chromosomal location for polymorphic gene families in the C. parvum genome. ( B ) Map of the MEDLE2 locus targeted in C. parvum for insertion of a 3× hemagglutinin (HA) epitope tag, a nanoluciferase reporter gene (Nluc), and neomycin phosphotransferase selection marker (Neo). ( C ) PCR mapping of the MEDLE2 locus using genomic DNA from wild type (WT) and transgenic (MEDLE2-HA) sporozoites, corresponding primer pairs are shown in ( B ), and thymidine kinase (TK) gene used as a control. Note the presence of two bands in the 5′–3′ amplification, indicating the presence of a transgene (3081 bp) and persistence of an unmodified copy (1174 bp), suggesting multiple copies of MEDLE2 in the C. parvum genome; also see . ( D, E ) HCT-8 cultures were infected with WYLE4-HA ( D ) or MEDLE2-HA ( E ) transgenic parasites and fixed after 24 hr for immunofluorescence assay (IFA). Red, antibody to HA; green, Vicia villosa lectin stain, VVL ; blue, Hoechst DNA dye. Additional genes targeted and the localizations of their products are summarized in and and .

Article Snippet: Other , Fluorescin Vicia villosa lectin stain , Vector Labs , Cat# FL-1231-2 , IF (1:1000).

Techniques: Selection, Marker, Transgenic Assay, Amplification, Infection, Immunofluorescence, Staining

Journal: eLife

Article Title: Evolutionary expansion of apical extracellular matrix is required for the elongation of cells in a novel structure

doi: 10.7554/eLife.55965

Figure Lengend Snippet: ( A–B ) in situ hybridization for dumpy mRNA in the lobed species D. melanogaster ( A ) and the non-lobed species D. biarmipes ( B ). Pink box outlines location of zoomed in images presented in A1 and B1. Relevant expression highlighted with arrow (purple/white) for strong expression, asterisk for weak expression, and arrowhead for clasper-specific expression. Expression observed in D. melanogaster at 44 hr APF is not present in all samples (see ). ( C–D ) aECM is labeled with Vicia villosa lectin (VVA; green) and apical membrane labeled with E-cadherin (Ecad; magenta) at 44 hr APF in D. melanogaster ( C ) and D. biarmipes ( D ). Location of respective cross sections indicated in yellow for lateral plate ( C2–D2 ) and blue for posterior lobe in D. melanogaster ( C1 ) and corresponding position in D. biarmipes ( D1 ). All cross-sections are oriented with apical side at the top and basal side at the bottom. White arrows highlight the crevice localization between the lateral plate and clasper, which the aECM fills in D. melanogaster ( C1 ), but only a weakly stained strand-like structure of aECM appears in D. biarmipes ( D1 ). Tendrils of aECM can also be observed connecting to the lateral plate in both species (red arrowheads). Relevant structures labeled: Posterior lobe (PL), lateral plate (LP), clasper (C), sheath (S), and phallus (P). Scale bar, 20 μm. n = at least five per experiment.

Article Snippet: The following primary antibodies were used: rat anti-alpha tubulin (tyrosinated) 1:500 (MAB 1864-I, MilliporeSigma), mouse anti-alpha tubulin (acetylated) 1:500 (T6793, Sigma-Aldrich), rat anti-Ecadherin 1:500 (DCAD2, DSHB), mouse anti-Fasciclin III 1:500 (7G10, DSHB), rabbit anti-histone H3 (phospho S10) 1:50 (ab5176, Abcam), goat anti-GFP 1:300 (ab6662, Abcam), fluorescein Vicia Villosa Lectin (VVA) 1:200 (FL-1231, Vector Laboratories).

Techniques: In Situ Hybridization, Expressing, Labeling, Staining

Journal: eLife

Article Title: Evolutionary expansion of apical extracellular matrix is required for the elongation of cells in a novel structure

doi: 10.7554/eLife.55965

Figure Lengend Snippet:

Article Snippet: The following primary antibodies were used: rat anti-alpha tubulin (tyrosinated) 1:500 (MAB 1864-I, MilliporeSigma), mouse anti-alpha tubulin (acetylated) 1:500 (T6793, Sigma-Aldrich), rat anti-Ecadherin 1:500 (DCAD2, DSHB), mouse anti-Fasciclin III 1:500 (7G10, DSHB), rabbit anti-histone H3 (phospho S10) 1:50 (ab5176, Abcam), goat anti-GFP 1:300 (ab6662, Abcam), fluorescein Vicia Villosa Lectin (VVA) 1:200 (FL-1231, Vector Laboratories).

Techniques: Plasmid Preparation, Construct, Recombinant, Sequencing, Software