α ncam 1  (R&D Systems)

Journal: bioRxiv

Article Title: Natural killer cells associate with epithelial cells in the pancreatic ductal adenocarcinoma tumor microenvironment

doi: 10.1101/2024.05.23.593868

Figure Lengend Snippet: A. Representative IMC pseudoimages displaying NK cell-epithelial cell co-localization (αSMA = blue; E-cadherin, pan-cytokeratin = pink; NKG2D = yellow, scale bar = 100 µm). B. Representative multiplex immunofluorescence image of human PDAC showing NK cell-epithelial cell co-localization (n = 6; pan-cytokeratin = pink, CD56 = yellow, NKG2D = cyan, scale bar = 50 µm). C. Cellular neighborhood analysis revealed 8 distinct cellular neighborhoods (red = high proportion of cells within cellular neighborhoods, blue = low proportion of cells within cellular neighborhoods). D. Cellular interaction analysis (blue = strong avoidance, red = strong interaction; FDR, P < 0.01) of IMC-defined cell subpopulations. E. Distance analysis per patient (n = 21) revealed that epithelial cells in PDAC are the closest to NK cells. Cell populations that were significantly distant from epithelial cells were calculated using 2-way ANOVA, Kruskal-Wallis test (* P < 0.05, *** P < 0.001, **** P < 0.0001;). F. Network graph of the top 75% of interactions between all IMC-defined cell populations in human PDAC showing epithelial cells and NK cells exist in similar networks.

Article Snippet: Samples were incubated with α-pan-cytokeratin-eFluor™ 660 (Invitrogen, CAT#: 50-9003-82) and α-CD56 (Invitrogen, CAT#: MA1-19129) antibodies overnight (25°C).

Techniques: Multiplex Assay, Immunofluorescence

Journal: bioRxiv

Article Title: Natural killer cells associate with epithelial cells in the pancreatic ductal adenocarcinoma tumor microenvironment

doi: 10.1101/2024.05.23.593868

Figure Lengend Snippet: A. Schematic for Transwell invasion assays. Average relative invasion + SEM of B. Donor NK #1 (n = 9), C. Donor NK #2 (n = 12), D. Donor NK #3 (n = 6), and E. Donor NK #4 (n = 10) upon CD44 neutralization. * P < 0.05 as determined by Wilcoxon matched-pairs signed rank test. F. Schematic of spheroid invasion assay. G. Representative 20X immunofluorescence images of outlined (yellow) PANC-1 spheroids (pan-cytokeratin; red) embedded with NK cells (CD56; green), pointed out with white arrows, treated with or without α-CD44. Scale bar = 250 µm. H. Zoomed inset of NK cells in the α-CD44 treatment group in G (white box). Average number of NK cells per PANC-1 spheroid + SEM from I. Donor NK #1 (- α-CD44, n = 42; + α-CD44, n = 53), J. Donor NK #2 (- α-CD44, n = 43; + α-CD44, n = 29), and K. Donor NK #3 (- α-CD44, n = 47; + α-CD44, n = 27); n = # of spheroids analyzed; * P < 0.05 as determined by Wilcoxon matched-pairs signed rank test.

Article Snippet: Samples were incubated with α-pan-cytokeratin-eFluor™ 660 (Invitrogen, CAT#: 50-9003-82) and α-CD56 (Invitrogen, CAT#: MA1-19129) antibodies overnight (25°C).

Techniques: Neutralization, Invasion Assay, Immunofluorescence

Journal: Glycobiology

Article Title: Site-specific immobilization of the endosialidase reveals QSOX2 is a novel polysialylated protein

doi: 10.1093/glycob/cwae026

Figure Lengend Snippet: Proteomic analysis of polysialylated proteins from NK-92 cells. A) Immunoblot analysis of NK-92 cell lysate samples treated with PBS or with active EndoN before isolation on immobilized EndoN DM . α-polySia and α-NCAM blots show lysate (L), flowthrough (F), wash (W), and boiled beads (B). B) Volcano plot of proteins identified in samples relative to those pretreated with active EndoN. Proteins with a significant difference are labelled. The known polysialylated proteins NCAM1 and ST8Sia4 are highlighted ( n = 4 per group, assessed with student’s T-test).

Article Snippet: The antibodies used in this manuscript consisted of α-polySia mAb 735 (BioAspect BA-Ab00240-2.0), α-CD56 (R&D systems, mAB2408), α-beta-tubulin (Abcam, ab6046), streptavidin-HRP (Biolegend, 405210), goat α-mouse IgG-HRP (R&D systems, HAF007), goat α-rabbit IgG-HRP (Abcam, ab6721), and α-QSOX2 (Abcam, ab191168 and ab121376).

Techniques: Western Blot, Isolation