Journal: Cellular and Molecular Life Sciences

Article Title: Defective IGF-1 prohormone N-glycosylation and reduced IGF-1 receptor signaling activation in congenital disorders of glycosylation

doi: 10.1007/s00018-022-04180-x

Figure Lengend Snippet: Lectin-binding analysis. Concanavalin A (ConA) and Phaseolus vulgaris leucoagglutinin (PHA-L) binding to CTR (mean of three technical replicates for each of five biological replicates) and CDG (mean of three technical replicates for each of twelve biological replicates) fibroblasts. Con A and PHA-L recognize high mannose and complex type N-glycans, respectively. *significantly different from CTR fibroblasts, * p < 0.0001

Article Snippet: For lectin blotting, membranes were probed with biotinylated Concanavalin A (ConA, 1:1000; Cat #B-1005-5) and Phaseolus vulgaris leucoagglutinin (PHA-L, 1:200; Cat #B-1115-2) lectins (Vector laboratories, D.B.A.

Techniques: Binding Assay

Journal:

Article Title: The Adhesin Hwp1 and the First Daughter Cell Localize to the a/a Portion of the Conjugation Bridge during Candida albicans Mating

doi: 10.1091/mbc.E03-04-0264

Figure Lengend Snippet: Multiple staining procedure. (A and B) DIC and FITC-ConA staining (green) of an a/a cell. (C and D) DIC and rhodamine-ConA staining (red) of an α/α cell. (E and F) DIC and calcofluor-staining of a/a or α/α cells. (G and H) DIC and anti-Hwp1 antibody-staining of an early conjugation tube formed by an α-pheromone-induced a/a cell. In the procedure, a/a and α/α parent cells are vitally stained with FITC-ConA and rhodamine-ConA, respectively, and then allowed to mate. The embryos are then fixed and stained with anti-Hwp1 antibody plus secondary antibody and calcofluor. Bar, 5 μm.

Article Snippet: To label the walls of living cells to differentiate between a / a and α/α parental cells in fusions, a / a cells were stained for 30 min at room temperature with rhodamine-concanavalin A (rhodamine-ConA) (Vector Laboratories, Burlingame, CA) and α/α cells with fluorescein isothiocyanate-concanavalin A (FITC-ConA) (Vector Laboratories), both diluted 1:100 from 5 mg/ml stock solutions, before mixing in mating experiments.

Techniques: Staining, Conjugation Assay

Journal:

Article Title: The Adhesin Hwp1 and the First Daughter Cell Localize to the a/a Portion of the Conjugation Bridge during Candida albicans Mating

doi: 10.1091/mbc.E03-04-0264

Figure Lengend Snippet: The multiple staining procedure demonstrates that in an a/a × α/α mating culture, only conjugation tubes formed by a/a cells contain Hwp1. FITC-ConA–stained (green) a/a cells and rhodamine-ConA–stained (red) α/α cells were mixed for 2 or 6 h and then fixed and stained with anti-Hwp1 antibody and calcofluor. (A–C; D–F) a/a cells with tubes. (G–I; J–L) α/α cells with tubes. +, staining of HWP1; –, no staining of Hwp1. Bar, 5 μm.

Article Snippet: To label the walls of living cells to differentiate between a / a and α/α parental cells in fusions, a / a cells were stained for 30 min at room temperature with rhodamine-concanavalin A (rhodamine-ConA) (Vector Laboratories, Burlingame, CA) and α/α cells with fluorescein isothiocyanate-concanavalin A (FITC-ConA) (Vector Laboratories), both diluted 1:100 from 5 mg/ml stock solutions, before mixing in mating experiments.

Techniques: Staining, Conjugation Assay

Journal:

Article Title: The Adhesin Hwp1 and the First Daughter Cell Localize to the a/a Portion of the Conjugation Bridge during Candida albicans Mating

doi: 10.1091/mbc.E03-04-0264

Figure Lengend Snippet: The multiple staining procedure demonstrates that only the a/a contribution to the conjugation bridge contains Hwp1. FITC-ConA–stained (green) a/a cells and rhodamine-ConA–stained (red) α/α cells were mixed for 18 h and then fixed and stained with anti-Hwp1 antibody and calcofluor. (A–C; D–F; G–I) Individual fusants before bud formation.. Bar, 5 μm.

Article Snippet: To label the walls of living cells to differentiate between a / a and α/α parental cells in fusions, a / a cells were stained for 30 min at room temperature with rhodamine-concanavalin A (rhodamine-ConA) (Vector Laboratories, Burlingame, CA) and α/α cells with fluorescein isothiocyanate-concanavalin A (FITC-ConA) (Vector Laboratories), both diluted 1:100 from 5 mg/ml stock solutions, before mixing in mating experiments.

Techniques: Staining, Conjugation Assay

Journal:

Article Title: The Adhesin Hwp1 and the First Daughter Cell Localize to the a/a Portion of the Conjugation Bridge during Candida albicans Mating

doi: 10.1091/mbc.E03-04-0264

Figure Lengend Snippet: The multiple staining procedure demonstrates that daughter cells form only on the a/a contribution to the bridge. FITC-ConA–stained (green) a/a cells and rhodamine-ConA–stained (red) α/α cells were mixed for 18 h and then fixed and stained with anti-Hwp1 antibody and calcofluor. (A–C; D–F; G–I; J–L) Individual zygotes. Bar, 5 μm.

Article Snippet: To label the walls of living cells to differentiate between a / a and α/α parental cells in fusions, a / a cells were stained for 30 min at room temperature with rhodamine-concanavalin A (rhodamine-ConA) (Vector Laboratories, Burlingame, CA) and α/α cells with fluorescein isothiocyanate-concanavalin A (FITC-ConA) (Vector Laboratories), both diluted 1:100 from 5 mg/ml stock solutions, before mixing in mating experiments.

Techniques: Staining

Journal:

Article Title: Identification of an Intestinal Neutral Glycosphingolipid as a Phenotype-Specific Receptor for the K88ad Fimbrial Adhesin of Escherichia coli †

doi:

Figure Lengend Snippet: Binding of K88ad adhesin to neutral glycosphingolipids from adhesive and nonadhesive phenotypes of pigs. Neutral glycosphingolipids (100 μg) from five phenotypes of pigs (A [lane 2], E [lane 3], B [lane 4], C [lane 5], and D [lane 6]) were separated on HPTLC plates as described in Materials and Methods. Chromatograms were either stained with orcinol-sulfuric acid reagent (panel I) or incubated with biotinylated K88ad adhesin (panel II) as described in Materials and Methods. Lane 1 contains glycolipid standards Lc2Cer (2), Gb4Cer (4), and Gb5Cer (5). The arrowheads indicate the positions of IGLad.

Article Snippet: Biotinylated vegetable lectins (concanavalin A lectin [ConA], soybean agglutinin [SBA], peanut agglutinin [PNA], Sambucus nigra agglutinin [SNA], Bandeira simplicifolia lectin [BSL II], Datura stramonium lectin [DSL], Jacalin, wheat germ agglutinin [WGA], Ulex europaeus agglutinin [UEA], Ricinus communis agglutinin [RCA 120 ], Maackia amurensis lectin I and II [MAL I and MAL II], Erythrina cristagalli lectin [ECL], Vicia villosa agglutinin [VVA]) were obtained from Vector Laboratories, Inc. (Burlingame, Calif.).

Techniques: Binding Assay, High Performance Thin Layer Chromatography, Staining, Incubation

Journal:

Article Title: Identification of an Intestinal Neutral Glycosphingolipid as a Phenotype-Specific Receptor for the K88ad Fimbrial Adhesin of Escherichia coli †

doi:

Figure Lengend Snippet: Determination of the binding of selected biotinylated lectins

Article Snippet: Biotinylated vegetable lectins (concanavalin A lectin [ConA], soybean agglutinin [SBA], peanut agglutinin [PNA], Sambucus nigra agglutinin [SNA], Bandeira simplicifolia lectin [BSL II], Datura stramonium lectin [DSL], Jacalin, wheat germ agglutinin [WGA], Ulex europaeus agglutinin [UEA], Ricinus communis agglutinin [RCA 120 ], Maackia amurensis lectin I and II [MAL I and MAL II], Erythrina cristagalli lectin [ECL], Vicia villosa agglutinin [VVA]) were obtained from Vector Laboratories, Inc. (Burlingame, Calif.).

Techniques: Binding Assay

Journal:

Article Title: Identification of an Intestinal Neutral Glycosphingolipid as a Phenotype-Specific Receptor for the K88ad Fimbrial Adhesin of Escherichia coli †

doi:

Figure Lengend Snippet: Binding of K88ad adhesin to intestinal neutral glycosphingolipids after β-galactosidase treatment. Neutral glycosphingolipids from a phenotype A animal were treated with β-galactosidase as described in Materials and Methods. Both treated (lane 3) and untreated (lane 2) glycosphingolipids (100 μg) were separated on HPTLC plates as described in Materials and Methods. Chromatograms were stained with orcinol-sulfuric acid reagent (A), or incubated with biotinylated K88ad adhesin (B) or RCA120 (C) as described in Materials and Methods. Lane 1 contains glycolipid standards Lc2Cer (2), Gb4Cer (4) and Gb5Cer (5). The arrowheads indicate the positions of IGLad.

Article Snippet: Biotinylated vegetable lectins (concanavalin A lectin [ConA], soybean agglutinin [SBA], peanut agglutinin [PNA], Sambucus nigra agglutinin [SNA], Bandeira simplicifolia lectin [BSL II], Datura stramonium lectin [DSL], Jacalin, wheat germ agglutinin [WGA], Ulex europaeus agglutinin [UEA], Ricinus communis agglutinin [RCA 120 ], Maackia amurensis lectin I and II [MAL I and MAL II], Erythrina cristagalli lectin [ECL], Vicia villosa agglutinin [VVA]) were obtained from Vector Laboratories, Inc. (Burlingame, Calif.).

Techniques: Binding Assay, High Performance Thin Layer Chromatography, Staining, Incubation

Journal:

Article Title: Identification of an Intestinal Neutral Glycosphingolipid as a Phenotype-Specific Receptor for the K88ad Fimbrial Adhesin of Escherichia coli †

doi:

Figure Lengend Snippet: Binding of K88ad adhesin to intestinal neutral glycosphingolipids after α-fucosidase treatment. Neutral glycosphingolipids from a phenotype A animal were treated with α-fucosidase as described in Materials and Methods. Both treated (lane 3) and untreated (lane 2) glycosphingolipids (100 μg) were separated on HPTLC plates as described in Materials and Methods. Chromatograms were stained with orcinol-sulfuric acid reagent (A) or incubated with biotinylated K88ad adhesin (B) or UEA (C) as described in Materials and Methods. Lane 1 contains glycolipid standards Lc2Cer (2), Gb4Cer (4) and Gb5Cer (5). The arrowheads indicate the positions of IGLad.

Article Snippet: Biotinylated vegetable lectins (concanavalin A lectin [ConA], soybean agglutinin [SBA], peanut agglutinin [PNA], Sambucus nigra agglutinin [SNA], Bandeira simplicifolia lectin [BSL II], Datura stramonium lectin [DSL], Jacalin, wheat germ agglutinin [WGA], Ulex europaeus agglutinin [UEA], Ricinus communis agglutinin [RCA 120 ], Maackia amurensis lectin I and II [MAL I and MAL II], Erythrina cristagalli lectin [ECL], Vicia villosa agglutinin [VVA]) were obtained from Vector Laboratories, Inc. (Burlingame, Calif.).

Techniques: Binding Assay, High Performance Thin Layer Chromatography, Staining, Incubation

Journal:

Article Title: Identification of an Intestinal Neutral Glycosphingolipid as a Phenotype-Specific Receptor for the K88ad Fimbrial Adhesin of Escherichia coli †

doi:

Figure Lengend Snippet: Binding of K88ad adhesin to purified neutral glycosphingolipids. Two micrograms of Lc3Cer (lane 1), Lc3Cer plus Lc4Cer (lane 2), nLc4Cer (lane 3), III4FucLc4Cer or Lea (lane 4), III3FucnLc4Cer or Lex (lane 5), V3FucnLc6Cer (lane 6), and VI2FucnLc6Cer (lane 7) were separated on HPTLC. These glycosphingolipids were either stained with orcinol-sulfuric acid reagent (A) or incubated with biotinylated K88ad adhesin (B) as described in Materials and Methods.

Article Snippet: Biotinylated vegetable lectins (concanavalin A lectin [ConA], soybean agglutinin [SBA], peanut agglutinin [PNA], Sambucus nigra agglutinin [SNA], Bandeira simplicifolia lectin [BSL II], Datura stramonium lectin [DSL], Jacalin, wheat germ agglutinin [WGA], Ulex europaeus agglutinin [UEA], Ricinus communis agglutinin [RCA 120 ], Maackia amurensis lectin I and II [MAL I and MAL II], Erythrina cristagalli lectin [ECL], Vicia villosa agglutinin [VVA]) were obtained from Vector Laboratories, Inc. (Burlingame, Calif.).

Techniques: Binding Assay, Purification, High Performance Thin Layer Chromatography, Staining, Incubation

Journal:

Article Title: Identification of an Intestinal Neutral Glycosphingolipid as a Phenotype-Specific Receptor for the K88ad Fimbrial Adhesin of Escherichia coli †

doi:

Figure Lengend Snippet: Comigration of IGLad and nLc4Cer on HPTLC plates. Neutral glycosphingolipids from a phenotype A animal (100 μg, lane 2), and nLc4Cer (2 μg, lane 3) were separated on HPTLC. These glycosphingolipids were either stained with orcinol-sulfuric acid reagent (A) or incubated with biotinylated K88ad adhesin (B) as described in Materials and Methods. Lane 1 contains glycolipid standards (Lc2Cer [2], Gb4Cer [4], and Gb5Cer [5]). The arrowheads indicate the position of IGLad.

Article Snippet: Biotinylated vegetable lectins (concanavalin A lectin [ConA], soybean agglutinin [SBA], peanut agglutinin [PNA], Sambucus nigra agglutinin [SNA], Bandeira simplicifolia lectin [BSL II], Datura stramonium lectin [DSL], Jacalin, wheat germ agglutinin [WGA], Ulex europaeus agglutinin [UEA], Ricinus communis agglutinin [RCA 120 ], Maackia amurensis lectin I and II [MAL I and MAL II], Erythrina cristagalli lectin [ECL], Vicia villosa agglutinin [VVA]) were obtained from Vector Laboratories, Inc. (Burlingame, Calif.).

Techniques: High Performance Thin Layer Chromatography, Staining, Incubation

Journal: Frontiers in Microbiology

Article Title: Roles of Three HSF Domain-Containing Proteins in Mediating Heat-Shock Protein Genes and Sustaining Asexual Cycle, Stress Tolerance, and Virulence in Beauveria bassiana

doi: 10.3389/fmicb.2018.01677

Figure Lengend Snippet: Contributions of Hsf1 , Sfl1 , and Skn7 to cell wall integrity of B. bassiana . (A) EC 50 for Congo red to suppress 50% colony growth of each strain after 6-day incubation at 25°C on 1/4 SDAY plates, to which small hyphal disks were attached for colony initiation. (B) Relative germination percentages of conidia after 24-h incubation at 25°C in a medium alone (control) or supplemented with Congo red (1 mg/mL). (C) Concentrations of protoplasts released from the hyphal cells after 6 h treatment with cell wall lysing enzymes in osmotic solution of 0.8 M sucrose. (D) Fluorescence intensity as an index of cell wall composition from flow cytometry of 2 × 10 4 conidia labeled with the fluorescent lectins ConA, GNL, and WGA. Asterisked bars in each bar group differ significantly from those unmarked (Tukey’s HSD, P < 0.05). Error bar: SD from three replicates.

Article Snippet: Carbohydrate epitopes on the surfaces of conidia were probed with the Alexa fluor 488-labeled lectins concanavalin A [ConA specific to α-glucose and α-N-acetylglucosamine (α-GlcNAc)], Galanthus nivalis (GNL lectin specific to mannose residues), and wheat germ agglutinin (WGA specific to β-GlcNAc and sialic acids) from Molecular Probes-Invitrogen and Vector Laboratories, respectively.

Techniques: Incubation, Fluorescence, Flow Cytometry, Labeling

Journal: Cell

Article Title: Small RNAs are modified with N-glycans and displayed on the surface of living cells

doi: 10.1016/j.cell.2021.04.023

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: All lectins were bought biotinylated from Vector labs: biotin-wheat germ agglutinin (WGA), biotin-concanavalin A (ConA), and biotin-Maackia Amurensis Lectin II (MAAII).

Techniques: Recombinant, Staining, Blocking Assay, Plasmid Preparation, High Performance Liquid Chromatography, Protein Extraction, Expressing, Software