Journal: bioRxiv

Article Title: Sialidases derived from Gardnerella vaginalis remodel the sperm glycocalyx and impair sperm function

doi: 10.1101/2025.02.01.636076

Figure Lengend Snippet: A) Schematic describing lectin flow cytometry and representative glycan epitopes. B, C, D) Sperm were exposed to sialidases (AUS 20 units or GvNanH2 160 units) for 1 hour, fixed in formalin, and stained with MAL-II-biotin/Neutravidin-Alexa Fluor 488, SNA-Cy5, or PSA-FITC respectively. Each point represents the median fluorescence intensity from one donor. Statistics represent a mixed model comparison with post-hoc Holm-Šidák tests. E) Schematic representing change in zeta potential after removal of negatively charged sialic acid on sperm surface. F) Sperm were treated with sialidase at various doses for 1 hour, and zeta potential was measured via dynamic light scattering. Each measurement represents the average of seven technical replicates, and statistics represent a mixed model comparison with post hoc Holm-Šidák tests.

Article Snippet: Biotinylated Maackia Amurensis Lectin II (MAL II, Cat #:B-1265-1) and Cy5-conjugated Sambucus Nigra Lectin (SNA, Cat #: CL-1305-1) were purchased from Vector Labs. Human Contraception Antibody (HCA), was a gift from ZabBio.

Techniques: Flow Cytometry, Staining, Fluorescence, Comparison, Zeta Potential Analyzer

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: MicroRNA-139 inhibits hepatocellular carcinoma cell growth through down-regulating karyopherin alpha 2

doi: 10.1186/s13046-019-1175-2

Figure Lengend Snippet: MiR-139 suppresses HCC migration and invasion. a - e Hep3B and SMMC7721 cells were transfected with 100 nM miR-139 mimic oligos or control oligos. The migration capacity of ( a ) Hep3B and ( b ) SMMC7721 cells were analyzed with the transwell migration assay. The invasion ability of ( c ) Hep3B and ( d ) SMMC7721 cells were measured with the matrigel transwell invasion assay. e The protein expression levels of E-cadherin, Vimentin and SNAIL1 were assayed with western blot in both Hep3B and SMMC7721 cells. f Hep3B and SMMC7721 cells were transfected with 100 nM miR-139 inhibitory or control oligos. The protein levels of E-cadherin, Vimentin and SNAIL1 were measured by western blot in both cell lines. All experiments were repeated 3 times. ** P < 0.01, *** P < 0.001

Article Snippet: The primary antibodies used in this study included: E-cadherin (1:1000, Abcam, USA), Vimentin (1:1000, Boster, China), SNAIL1 (1:1000, Boster, China), KPNA2 (1:1000, Abcam, USA), c-myc (1:1000, Beyotime, China), POU5F1 (1:1000, Abcam, USA), and β-actin (1:2000, Abcam, USA).

Techniques: Migration, Transfection, Control, Transwell Migration Assay, Transwell Invasion Assay, Expressing, Western Blot

Journal: Cancers

Article Title: Decoding Single Cell Morphology in Osteotropic Breast Cancer Cells for Dissecting Their Migratory, Molecular and Biophysical Heterogeneity

doi: 10.3390/cancers14030603

Figure Lengend Snippet: Differential binding of lectins to breast cancer cell lines. ( A , B ) The parental cells MB-231 and its bone-seeking derivatives, MET and BONE, were cell-surface labeled with one of a panel of distinct FITC-conjugated lectins prior to cytochemistry (( A ), left panels) and flow cytometry (( A ), right panels, ( B )) analyses. See also . Percentages of positive cells are indicated in the histograms ( A ) and the median fluorescence intensity is presented for each lectin ( B ). Data from representative experiments acquired under uniform instruments setting for all cell lines are displayed. The lectins used were Concanavalin A (ConA), Datura Stramonium Lectin (DSL), Dolichos Biflorus Agglutinin (DBA), Erythrina Cristagalli Lectin (ECL), Griffonia Simplicifolia Lectin I (GSL-I), Griffonia Simplicifolia Lectin II (GSL-II), Lens Culinaris Agglutinin (LCA), Lycopersicon Esculentum Lectin (LEL), Peanut Agglutinin (PNA), Phaseolus Vulgaris Lectin E (PHA-E), Phaseolus Vulgaris Lectin L (PHA-L), Pisum Sativum Agglutinin (PSA), Ricinus Communis Agglutinin I (RCA), Sambucus Nigra Lectin (SNA), Solanum Tuberosum Lectin (STL), Soybean Agglutinin (SBA), Ulex Europaeus Agglutinin I (UEA-I), Vicia Villosa Lectin (VVL) and Wheat Germ Agglutinin (WGA, succinylated (succ)-WGA). Scale bar, 25 μm.

Article Snippet: After inactivation of trypsin, 2 washing steps with PBS and centrifugation (5 min at 300× g ), cells were resuspended in PBS or Ca/Mg-PBS (for lectin labeling), containing 1% BSA and 100 μL-cell suspension aliquots were incubated with unconjugated or fluorochrome-conjugated primary antibodies ( ) or FITC- or biotin-conjugated lectins (see above, Lectin kit I, Biotinylated (BK-1000) and SNA, Biotinylated (B-1305-2), both from Vector Laboratories) for 30 min at 4 °C.

Techniques: Binding Assay, Labeling, Flow Cytometry, Fluorescence