dolichos biflorus agglutinin dba  (Vector Laboratories)

Journal: Scientific Reports

Article Title: Effects of exercise on vascular remodelling and fetal growth in uncomplicated and abortion-prone mouse pregnancies

doi: 10.1038/s41598-024-83329-z

Figure Lengend Snippet: Impact of exercise and paternal genotype on the proportion of uterine natural killer cells in the decidua at mid-pregnancy. DBA lectin-stained sections show decidual areas of sedentary ( A , E , C , G ) and exercised ( B , F , D , H ) mice. Quantification of the DBA + uNK cells are identified ( I ) within the total decidual area ( J ) for sedentary (S) and exercised (E) CBA/J x BALB/c mice (circles, A , B , E , F ) and abortion-prone CBA/J x DBA/2J mice (triangles, C , D , G , H ) in mid-pregnancy ( N = 5–18 dams/group). Data was analysed by linear mixed model including litter size as a covariate. Symbols show outcomes from each dam and bars and whiskers indicate mean ± SD within each group. Differences between male genotypes are indicated by * P < 0.05; ** P < 0.01. Arrowheads define DBA + uNK cells.

Article Snippet: To visualise uNK cells within GD10.5 implantation sites, tissue sections were stained using Dolichos Biflorus Agglutinin (DBA) lectin (Vector Laboratories), counterstained with Weigert’s haematoxylin .

Techniques: Staining

Journal: bioRxiv

Article Title: Ductal pancreatic cancer interception by FGFR2 abrogation

doi: 10.1101/2024.10.16.618726

Figure Lengend Snippet: A. Left, representative IF for the acinar marker AMY (white), the ductal marker Dolichos Biflorus Agglutinin (DBA) (green), FGFR2 (red) and 4′,6-diamidino-2-phenylindole (DAPI, blue) conducted on healthy pancreata from B6J mice (n = 5) and ADM and mPanINs from KC mice (n = 6). Scale bars, 25 μm. Right, quantification of staining plotted as mean ± SD. 3 images per mouse were quantified. Unpaired Student’s t test. B. Left, representative IF for CK19 (white), FGFR2 (red) and DAPI (blue) conducted on mPanINs from KC mice (n = 7). Scale bars, 25 μm. Right, quantification of staining plotted as mean ± SD. 3 images per mouse were quantified. C. Left, representative IF for CK19 (white), p53 (green), FGFR2 (red) and DAPI (blue) conducted on tumors from KPC mice (n = 6). Scale bars, 25 μm. Right, quantification of staining plotted as mean ± SD. 3 images per mouse were quantified. Unpaired Student’s t test. D. Representative IHC for FGFR2 conducted on human IPMN and PanIN lesions. E. Left, representative IF for FGFR2 (red), GATA6 (white), S100A2 (green) and DAPI (blue) conducted on human PDAs. Scale bars, 25 μm. Samples that present % GATA6 + S100A2 - cancerous cells > 60 are classified as ‘Classical’, samples with % GATA6 - S100A2 + cancerous cells > 30 are classified as ‘Basal-like’ and samples with % GATA6 + S100A2 + cancerous cells > 5 are classified as ‘IC’. Remaining samples that don’t meet these criteria are classified as ‘other’. Right, quantification of staining plotted as mean ± SD. Unpaired Student’s t test. F. Schematic representation of FGFR2 expression during pancreatic cancer progression in the KRAS G12D -driven mouse model and in human samples.

Article Snippet: To perform IHC, endogenous peroxidase activity was quenched in 3% H 2 O 2 for 20 min. Tissues were blocked in 2.5% Normal Horse Serum blocking solution (Vector Laboratories) for IHC or 5% BSA (Sigma) in TBST buffer for IF and subjected to staining with the following antibodies overnight at 4C: p53 (Leica, P53-CM5P-L 1:100), GFP/YFP (Abcam, ab6673 1:100), AMYLASE (Sigma, A8273 1:250), Dolichos Biflorus Agglutinin (DBA) Fluorescein (VectorLabs, FL-1031 1:200), FGFR2 (Abcam, ab58201 1:1000), CK19 Alexa Fluor 647 (Abcam, ab205446 clone EPR1579Y 1:500), CK19 (Sigma, MABT913 1:500), GATA6 (R&D, AF1700 1:500), S100A2 (Abcam, ab109494 1:250), FGF7 (Origene, TA321423 1:100), FGF10 (Sigma, ABN44 1:100), phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (CST, 4370 clone D13.14.4E 1:250), Ki67 (ThermoFisher, 14-5698-82 clone SolA15 1:500), Ki67 Alexa Fluor 488 (Abcam, ab281847 clone SP6 1:500), clusterin (ThermoFisher, PA5-46931 1:100).

Techniques: Marker, Staining, Expressing

Journal: mBio

Article Title: Constitutive upregulation of transcription factors underlies permissive bradyzoite differentiation in a natural isolate of Toxoplasma gondii

doi: 10.1128/mbio.00641-24

Figure Lengend Snippet: In vitro phenotypes and genetic characterization of Tg68. ( A ) HFF cells were infected with Tg68 or ME49 tachyzoites at an MOI of 5 for 24, 48, or 72 h. Cells were fixed and IF staining was performed with anti-SAG1 rabbit antibody and goat anti-rabbit secondary antibody conjugated to Alexa Fluor 647. Hoechst 33342 was used to stain DNA. Vacuoles were visualized using a Cytation 3 Imager and representative images are shown. Scale bar, 50 µm. ( B ) Violin plot representing diameters of vacuoles of Tg68 or ME49 tachyzoites at indicated timepoints. Data are combined from three independent experiments in which at least 5,000 vacuoles per strain were measured in each experiment. In violin plots, the middle line indicates median, the upper line indicates the 75th percentile, and the lower line indicates the 25th percentile. ***Student’s t -test, P < 0.001. ( C ) HFF cells infected with Tg68 or ME49 tachyzoites were cultured under alkaline pH conditions (pH = 8.2, ambient CO 2 ) for 7 days. Cells were fixed and stained with anti-BAG1 mouse mAb 8.25.8 and anti-SAG1 rabbit antibody, followed by goat anti-mouse secondary antibody conjugated to Alexa Fluor 568 and goat anti-rabbit secondary antibody conjugated to Alexa Fluor 647. Dolichos biflorus agglutinin (DBA) conjugated to Alexa Fluor 488 was used to stain the cyst wall and Hoechst 33342 was used to stain DNA. Scale bar, 20 µm. ( D ) HFF cells infected with SAG1-EGFP BAG1-mCherry Tg68 and SAG1-EGFP BAG1-mCherry ME49 tachyzoites were cultured under alkaline pH conditions (pH = 8.2, ambient CO 2 ) for 7 days. Cells were fixed, and DBA conjugated to Alexa Fluor 488 was used to stain the cyst wall. Average percentages of BAG1-positive, BAG1-positive and SAG1-positive, and SAG1-positive parasites within individual DBA-positive cysts were determined using CellProfiler. Mean ± SE plotted for 3 independent experiments in which a minimum of 39 cysts and a maximum of 84 cysts were quantified from 15 separate fields of view for each strain in each experiment. *Student’s t -test, P < 0.05; *** P < 0.001. ( E ) Neighbor-net analysis of Tg68 and 62 T. gondii isolates based on 804,624 SNPs. Major clades of T. gondii are indicated in different colors. Scale bar, number of SNPs per site.

Article Snippet: Fluorescein-conjugated Dolichos Biflorus Agglutinin (DBA) was obtained from Vector Laboratories.

Techniques: In Vitro, Infection, Staining, Cell Culture