lens culinaris agglutinin  (Vector Laboratories)

Journal: Cell metabolism

Article Title: Cytoarchitectural multi-depot profiling reveals immune-metabolic crosstalk in human colon-associated adipose tissue

doi: 10.1016/j.cmet.2025.12.008

Figure Lengend Snippet: (A) Uniform manifold approximation and projection displaying adipocyte identities and expression densities for main marker genes in the STx data. (B) Dot plots displaying the scaled average expression (expr.) profiles of the top three marker genes for each adipocyte identity in the present work and a previous study. (C) Abundance (abun.) and depot enrichments (scaled proportions, prop.) for the three main adipocyte identities. Significant enrichments ( p < 0.05 using likelihood ratio test, LRT) are indicated. (D) Heatmaps displaying expressional enrichments of Adipo SAA markers across depots. Results are shown as scaled average expression for bulk transcriptomics and proteomics from Krieg et al., as well as STx and snRNA-seq in the main (1) and validation (2) cohorts. (E) Quantification of CES1 and lectin (LCA) normalized mean fluorescence intensity based on tissue sections from six individuals imaged using confocal microscopy. Asterisks denote statistically significant differences (* p < 0.05) comparing depots using one-way repeated measures ANOVA and Tukey’s multiple comparisons test. Error bars are standard error of the mean. (F) Representative image of epiploic WAT stained with LCA and an antibody directed against CES1. Scale bars, 200 μm (merge) and 100 μm (inlay). epi, epiploic; mc, mesocolic; ms, mesenteric; om, omental; sc, subcutaneous.

Article Snippet: Fluorescein-labeled lectin (1:200, FL-1041, Vector labs) or Dylight649-labeled lectin (1:200, DL-1048–1, Vector labs), together with Hoechst33342 (1:2000, ab228551, Abcam), was applied on the slides for 20 minutes.

Techniques: Expressing, Marker, Transcriptomics, Biomarker Discovery, Fluorescence, Confocal Microscopy, Staining

Journal: Cancer Medicine

Article Title: FUT8 Is a Critical Driver of Prostate Tumour Growth and Can Be Targeted Using Fucosylation Inhibitors

doi: 10.1002/cam4.70959

Figure Lengend Snippet: Targeting FUT8‐mediated core fucosylation in prostate cancer with fucosylation inhibitors suppresses tumour growth. (A) CWR22Rv1 cells were subcutaneously injected into the flank of 7‐week‐old CD‐1 nude mice. 7 days prior to implantations mice were randomised to start treatment with either 150 mg/kg fucosylation inhibitor SGN‐2FF or water via oral gavage daily ( n = 10 mice/group). Tumour size was measured every 3–4 days using callipers. (B) Tumour volume (mm 3 ) was significantly reduced in the SGN‐2FF treated mice after 21 days (Welch's t ‐test for tumour volume on Day 21, p = 0.0034, **). Representative images of tumours are shown. (C) Analysis of CWR22Rv1 xenograft tumours (from experiment shown in Figure 6B) using N ‐glycan MALDI‐MSI to identify core‐fucosylated N ‐glycans. Images show the spatial distribution of core fucosylated bi‐antennary N ‐glycan (1773.581 m/z), tri‐antennary N ‐glycan (1825.5961 m/z) and the complex core fucosylated tetra‐antennary N‐ glycan (2190.7632 m/z). EndoF3 cleavage induced a shift of 349.137 amu. Glycan nomenclature: blue square indicates GlcNAc, yellow circle indicates galactose, green circle indicates mannose, red triangle indicates fucose, and purple diamond indicates sialic acid. Scale bar is 5 mm. (D) WST‐1 cell proliferation assays show FUT8 overexpression significantly increases the proliferation of CWR22RV1 cells (unpaired t ‐test, p < 0.0001, ****), and this is suppressed by treatment with 30 μM of Fucotrim I over 72 h (unpaired t ‐test, p < 0.0001, ****). WST‐1 cell proliferation assays also show FUT8 knockdown significantly reduces the proliferation of PC3 cells (unpaired t ‐test, p < 0.0001, ****) and by treatment with 30 μM of Fucotrim I for 72 h (unpaired t ‐test, p = 0.0012, ***). (E) Colony formation assays show FUT8 overexpression significantly increases the ability of CWR22RV1 cells to survive and grow in colonies over 14 days (unpaired t ‐test, p < 0.0001, ****), and this is suppressed by treatment with 30 μM Fucotrim I (unpaired t test, p < 0.0001, ****). PC3 cells with knockdown of FUT8 have reduced colony formation over 14 days (unpaired t ‐test, p = 0.019, **). PC3 cells treated with 30 μM Fucotrim I for 14 days have reduced ability to survive and grow in colonies over 14 days (unpaired t ‐test, p = 0.0015, ***). (F) Inhibition of fucosylation in TRAMPC2 and RM1 mouse prostate cancer cells Fucotrim I detected using LCA lectin flow cytometry (which recognises core fucosylated N‐glycans . Cells were treated with a range of concentrations of Fucotrim I from 1 nM to 128 μM for 72 h. The mean fluorescence intensities were normalized to a DMSO control. (G) Colony formation assays show treatment with 64 μM Fucotrim significantly reduced cell colony formation for both TRAMPC2 cells (unpaired t ‐test, p < 0.0001, ****) and RM1 cells (unpaired t ‐test, p < 0.0001, ****) over 7 days.

Article Snippet: The cells were washed twice with 1X Carbo‐Free Blocking Solution (1X CFB) (Vector labs, SP‐5040‐125) then resuspended in 100 μL of 1:2000 FITC‐conjugated AAL lectin (Vector Labs, FL‐1391‐1) or LCA lectin (Vector Labs, FL‐1041‐5) in 1X CFB and incubated for 30 min at 4°C.

Techniques: Injection, Glycoproteomics, Over Expression, Knockdown, Inhibition, Flow Cytometry, Fluorescence, Control