fitc sna fl 1301  (Vector Laboratories)

Journal: bioRxiv

Article Title: 2-D organoids demonstrate specificity in the interactions of parasitic nematodes and their secreted products at the basal or apical intestinal epithelium

doi: 10.1101/2024.12.16.628471

Figure Lengend Snippet: A) Schematic of experimental culture conditions for the generation of 2-D enteroid cultures in Matrigel-coated transwell plates. B) Representative fluorescence microscopy image showing complete transwell coverage by 2-D enteroid monolayer stained with 4’,6-diamindino-2-phenylindole (DAPI) for nuclei (blue). C) Representative confocal immunofluorescence image showing organisation of Ki67+ Alexa Fluor 647 (AF647, yellow) cells into proliferative loci, surrounded by terminally differentiated goblet cells labelled with UEA & SNA FITC (green). Nuclei are stained with DAPI (cyan) and F-actin labelled with phalloidin (Magenta). D) Z-stack projection of confocal immunofluorescence image demonstrating apical brush border of epithelial cells by co-staining of microvilli marker Villin AF647 (Magenta) and F-actin using Phallodin AF594 (White). Ulex europaeus agglutinin I (UEA) fluorescein (FITC) and Sambucus nigra Lectin (SNA) FITC binds lectins present in mucus of goblet cells, UEA & SNA FITC (green) can be seen emerging through the brush border. E-I) Representative fluorescence microscopy images of 2-D enteroids with DAPI-stained nuclei, phalloidin AF594 binding F-actin, and UEA/SNA labelling goblet cells, and antibody-based immunohistochemistry in AF647 of E) villin on microvilli of enterocytes, F) the Ki67+ stem and transit-amplifying cells, G) lysozyme 1-expressing Paneth cells, H) Dckl1 expression on tuft cells and I) chromogranin A on enteroendocrine cells.

Article Snippet: Permaebilisation/block buffer was removed and samples were incubated with primary antibodies α-Lysozyme 1 (1:40, DAKO, A0099), α-DCAMKL1 (Abcam, UK, ab31704), α-Villin (1:100, Abcam, ab130751), α-Chromogranin-A (1:50, Abcam, ab15160), α-Ki67 (1:250, Abcam, ab16667) and the lectin dyes Sambucus Nigra FITC (SNA 1:50, Vector laboratories, UK, FL-1301) and Ulex europaeus FITC (UEA 1:1000, Sigma-aldrich, 19337) in diluent (PBS, 5% BSA, 0.25% Triton-X) overnight at 4°C.

Techniques: Fluorescence, Microscopy, Staining, Immunofluorescence, Marker, Binding Assay, Immunohistochemistry, Expressing

Journal: Cancers

Article Title: Sialylation Inhibition Can Partially Revert Acquired Resistance to Enzalutamide in Prostate Cancer Cells

doi: 10.3390/cancers16172953

Figure Lengend Snippet: Enzalutamide-resistant prostate cancer cells have upregulation of ST6GAL1 and increased levels of α2,6-sialylated glycans. ( A ) SNA lectin immunofluorescence shows VCaP EnzR cells have increased levels of ST6GAL1 and α2-6 sialylation (SNA, the lectin from Sambucus nigra, recognises α2-6-linked sialylated N-glycans ). ( B ) SNA lectin immunofluorescence shows LNCaP EnzR cells have increased levels of ST6GAL1 and α2-6 sialylation compared to control LNCaP cells. DNA is stained with Hoechst. Scale bar = 200 µM.

Article Snippet: The cells were incubated overnight at 4 °C with FITC-conjugated SNA lectin (Vector Laboratoriesabs, FL-1301-2).

Techniques: Immunofluorescence, Control, Staining

Journal: Cancers

Article Title: Sialylation Inhibition Can Partially Revert Acquired Resistance to Enzalutamide in Prostate Cancer Cells

doi: 10.3390/cancers16172953

Figure Lengend Snippet: The sialyltransferase inhibitor P-SiaFNEtoc blocks α2,6 sialylation in the VCaP EnzR and LNCaP EnzR prostate cancer cells. ( A ) Detection of immunofluorescent staining of ST6GAL1 and α2,6-sialylation of N -glycans in the VCaP control and VCaP EnzR cells treated with 20 µM of the sialyltransferase inhibitor P-SiaFNEtoc for 6 days. Treatment of both cell lines with P-SiaFNEtoc inhibits α2,6-sialylation of N -glycans (detected using SNA lectin). Control cells were treated with DMSO. Scale bar = 50 µm. The images are representative of three biological repeats. ( B ) Detection of immunofluorescent staining of ST6GAL1 and α2,6-sialylation of N -glycans in LNCaP control and LNCaP EnzR cells treated with 2 µM of the sialyltransferase inhibitor P-SiaFNEtoc for 3 days. Treatment of both cell lines with P-SiaFNEtoc inhibits α2,6-sialylation of N -glycans (detected using SNA lectin). The control cells were treated with DMSO. Scale bar = 50 µm. The images are representative of three biological repeats.

Article Snippet: The cells were incubated overnight at 4 °C with FITC-conjugated SNA lectin (Vector Laboratoriesabs, FL-1301-2).

Techniques: Staining, Control