Journal: Scientific Reports

Article Title: Targeting cell-derived markers to improve the detection of invisible biological traces for the purpose of genetic-based criminal identification

doi: 10.1038/s41598-023-45366-y

Figure Lengend Snippet: The principal carbohydrate patterns recognized by the 4 lectins tested.

Article Snippet: Samples were incubated for 20 min in a humidity chamber using a biotinylated lectin solution (at final concentration 10 µg/ml): Concanavalin A (ConA), Peanut agglutinin (PNA), Elderberry lectin (SNA), and Ulex europaeus agglutinin (UEA) (B-1005, B-1075, B-1305, and B-1065 respectively, Vector Laboratories, USA).

Techniques:

Journal: PLoS Pathogens

Article Title: GM1 ganglioside-independent intoxication by Cholera toxin

doi: 10.1371/journal.ppat.1006862

Figure Lengend Snippet: (A) Representative histograms from flow cytometry analysis of CTB- (red) and OVA- (white) binding to cell types in human peripheral blood. (B) Bar graphs show the geometric mean fluorescent index (gMFI) of CTB binding (with OVA gMFI subtracted) to human whole blood (n = 8) and murine splenocytes (n = 4–6). Each dot represents one donor/mouse. (C) gMFI of CTB and OVA binding to the different cell types from human whole blood. Each pair of dots represents one donor. (D-E) Histogram and bar graph (n = 8) showing blocking of CTB binding to human granulocytes by pre-treating CTB with (D) L-, D-fucose and D-galactose or (E) with the lectins AAL or MAL-II. (F) Histograms and bar graphs (n = 6) showing the blocking of CTB binding to murine (wt or KO) cells by either pre-treating the cells with lectins or pre-treating CTB with sugars. For panel (C) significance was calculated using paired t-test and for (D-E) one-way-ANOVA was used with Tukey correction (*** = p<0,005, **** = p<0,0001).

Article Snippet: Cells were then incubated for 30 min on ice with diluted biotinylated CTB (Thermo Fisher Scientific/Invitrogen, Cat. No. C34779) or biotinylated lectins (Vector Labs: PNA, Cat. B-1075; MAL-II, Cat. No. B-1265; AAL, Cat. No. B-1395) at the following concentrations: 100 μl of 10 μg/ml was added per well.

Techniques: Flow Cytometry, Binding Assay, Blocking Assay

Journal: PLoS Pathogens

Article Title: GM1 ganglioside-independent intoxication by Cholera toxin

doi: 10.1371/journal.ppat.1006862

Figure Lengend Snippet: (A) Bar graph showing relative absorbance values from an ELISA with immobilized anti-Le X , and detection with CTB-HRP. Samples as indicated from lysates of isolated human cells (2 μg protein/ml). Each dot represents a human donor (n = 5–8). (B) CD66 or (C) CD66 and Le X expression by jejunal epithelial cells that were isolated using EDTA medium (villi) or enzymatic degradation after EDTA treatment (non-villi or crypt). Histograms from flow cytometry analyses of CTB-, G33D- and OVA-binding to the differentially enriched epithelial cells. (B) EpCAM + cells and (C) EpCAM + Le X+ cells. (D-G) Bar graph showing percent of gMFI of CTB binding to jejunal epithelial cells by pretreatment of the cells with (D) lectins, (E) sugars, (F) oligosaccharides and (G) HSA-linked oligosaccharides. Graphs show the percent of gMFI of CTB binding to the cells where 100% represents CTB staining with no blocking oligosaccharide. Each dot represents a donor in (D) n = 4–12, (E) n = 6–8, (F) n = 6–12, (G) n = 6–7. Significance was calculated using a one-way-ANOVA with Tukey correction compared to CTB without block if not indicated otherwise with bars (**** = p<0,0001, *** = p<0,005, ** = p<0,01 and * = p<0,05).

Article Snippet: Cells were then incubated for 30 min on ice with diluted biotinylated CTB (Thermo Fisher Scientific/Invitrogen, Cat. No. C34779) or biotinylated lectins (Vector Labs: PNA, Cat. B-1075; MAL-II, Cat. No. B-1265; AAL, Cat. No. B-1395) at the following concentrations: 100 μl of 10 μg/ml was added per well.

Techniques: Enzyme-linked Immunosorbent Assay, Isolation, Expressing, Flow Cytometry, Binding Assay, Staining, Blocking Assay

Journal: PLoS Pathogens

Article Title: GM1 ganglioside-independent intoxication by Cholera toxin

doi: 10.1371/journal.ppat.1006862

Figure Lengend Snippet: (A-E) C6 cells were cultured with the indicated inhibitors for 72 h followed by: (A) Staining was then performed with biotin-CTB, followed by DTAF-streptavidin. Fluorescence was measured by flow cytometry, represented here by MFI. (B) 1 h exposure to CT after which accumulated cAMP was measured by the cAMP-Glo™ luminescence assay. Luminescence signal is inversely proportional to cAMP levels. (C) As in panel A , but stained with biotin-PNA, followed by DTAF-streptavidin (D) Cell lysates were separated by PAGE and probed with biotin-PNA, biotin-CTB, or no biotinylated reagent, followed by streptavidin-peroxidase conjugate and development with chemiluminescent substrate. Equivalent amounts of protein were loaded in each lane and blots were probed with an anti-α-tubulin or anti-GAPDH antibody to confirm equivalent loading. (E) As in panel B , but brefeldin A (BFA) was added 1 h prior to CT addition and was also present during CT induction.

Article Snippet: Cells were then incubated for 30 min on ice with diluted biotinylated CTB (Thermo Fisher Scientific/Invitrogen, Cat. No. C34779) or biotinylated lectins (Vector Labs: PNA, Cat. B-1075; MAL-II, Cat. No. B-1265; AAL, Cat. No. B-1395) at the following concentrations: 100 μl of 10 μg/ml was added per well.

Techniques: Cell Culture, Staining, Fluorescence, Flow Cytometry, Luminescence Assay

Journal: PLoS Pathogens

Article Title: GM1 ganglioside-independent intoxication by Cholera toxin

doi: 10.1371/journal.ppat.1006862

Figure Lengend Snippet: (A) Representative histograms from flow cytometry analyses of CTB-, G33D- and OVA-binding to WT and KO jejunal cells (non-villi epithelial and CD45+ cells). (B) Bar graph showing CTB, G33D and OVA gMFI of non-villi jejunal epithelial cells from wt (black) and KO (gray). (C-F) Bar graphs showing percent of gMFI of non-blocked CTB binding to non-villi jejunal epithelial cells (wt black and KO gray) following pretreatment of the cells with (C) lectins or CTB with (D) sugars, (E) oligosaccharides and (F) HSA-linked oligosaccharides. Graphs show the percent of gMFI of CTB binding to the cells where 100% represents CTB staining with no blocking. (G-H) Bar graph showing intestine-animal ratio (by weight) for WT and KO mice gavaged with PBS with or without CT. (H) KO mice were fed chow with (open circles) or without (full circles) NB-DNJ for 4 weeks prior to gavage. The graphs are from pooled experiments where each dot represents one animal (n = 4–30). Significance was calculated using a one-way-ANOVA with Tukey correction (**** = p<0,0001, *** = p<0,005, ** = p<0,01 and * = p<0,05).

Article Snippet: Cells were then incubated for 30 min on ice with diluted biotinylated CTB (Thermo Fisher Scientific/Invitrogen, Cat. No. C34779) or biotinylated lectins (Vector Labs: PNA, Cat. B-1075; MAL-II, Cat. No. B-1265; AAL, Cat. No. B-1395) at the following concentrations: 100 μl of 10 μg/ml was added per well.

Techniques: Flow Cytometry, Binding Assay, Staining, Blocking Assay

Journal: The Journal of Neuroscience

Article Title: Proteoglycan IMPG2 Shapes the Interphotoreceptor Matrix and Modulates Vision

doi: 10.1523/JNEUROSCI.2994-19.2020

Figure Lengend Snippet: List of names and specifications of antibodies and lectins used in this work

Article Snippet: PNA , , N -acetylgalactosamine , 1:1000 , Vector Laboratories , B-1075.

Techniques: Plasmid Preparation