fitc conjugated maa i  (Vector Laboratories)

Journal: Journal of Virology

Article Title: Modulation of cytokeratin and cytokine/chemokine expression following influenza virus infection of differentiated human tonsillar epithelial cells

doi: 10.1128/jvi.01460-24

Figure Lengend Snippet: Distribution of CK and lectin staining in HTECs as determined by flow cytometry. ( A ) HTECs were fixed and permeabilized, then stained with monoclonal antibodies specific for CK5, CK14, CK8/18, and CK19 for flow cytometric analysis. Percentages give fraction positive for the indicated CK when gated against matched isotype controls. ( B ) HTECs were stained with MAA I-FITC lectin for avian-adapted α2,3-linked SA receptors or with SNA-BV786 lectin for mammalian-adapted α2,6-linked SA receptors. (B.a) MAA I lectin staining was nearly universal in more than five replicate experiments. (B.b) SNA lectin staining was more limited over the same experiments. (B.c) Dual staining showed that coincident expression of avian with mammalian-adapted SA in HTECs was robust. Gating is based on unstained and fluorescence-minus-one controls as shown in . Inset statistics give means ± SEM of the indicated populations from three replicate experiments.

Article Snippet: Uninfected HTECs were first stained with biotinylated SNA lectin SA and MAA I FITC-conjugated lectin (Vector Laboratories).

Techniques: Staining, Flow Cytometry, Expressing, Fluorescence

Journal: eBioMedicine

Article Title: Sialic acid blockade inhibits the metastatic spread of prostate cancer to bone

doi: 10.1016/j.ebiom.2024.105163

Figure Lengend Snippet: Sialic acid blockade can prevent/inhibit prostate cancer bone metastasis. ( a ) Inhibition of sialylation in TRAMPC2 cells using P-SiaFNEtoc detected using pan-specific Lectenz lectin flow cytometry. Cells were treated with a range of concentrations of P-SiaFNEtoc inhibitor from 2 μM to 512 μM for 72 h. The intensities were normalised to a DMSO control. ( b ) Detection of α2-6 linked sialylated N -glycans in TRAMPC2 cells using SNA lectin flow cytometry. TRAMPC2 cells treated with 64 μM P-SiaFNEtoc for 72 h had reduced levels of SNA binding indicating a reduction in α2-6 linked sialylation in these cells (unpaired t test, p = 0.0001). ( c ) Luciferase tagged TRAMPC2 cells (control or pre-treated with 64 μM P-SiaFNEtoc for 72 h) were injected into immunocompetent C57BL/6 mice via sub-cutaneous injection and tumours were monitored using in vivo bioluminescence imaging. Pre-treatment of TRAMPC2 cells with P-SiaFNEtoc (which removed sialylated glycans) significantly reduced tumour burden over 6 weeks (n = 10, Mann–Whitney test, p = 0.0233) thus suggesting that sialic acid blockade has the potential to inhibit the growth of prostate tumours. ( d ) Inhibition of sialylation in RM1 cells using P-SiaFNEtoc detected using pan-specific Lectenz lectin flow cytometry. Cells were treated with a range of concentrations of P-SiaFNEtoc inhibitor from 2 μM to 512 μM for 72 h. The intensities were normalised to a DMSO control. ( e ) Detection of α2-6 linked sialylated N -glycans in RM1 cells using SNA lectin flow cytometry. RM1 cells treated with 256 μM P-SiaFNEtoc for 72 h had reduced levels of SNA binding indicating a reduction in α2-6 linked sialylation in these cells (unpaired t test, p < 0.0001). ( f ) Luciferase tagged RM1 cells (control or pre-treated with 256 μM P-SiaFNEtoc for 72 h) were injected into immunocompetent C57BL/6 mice via intra cardiac injection. Tumours were monitored over 15 days using in vivo bioluminescence imaging. ( g , h ) Pre-treatment of RM1 cells with P-SiaFNEtoc (to remove sialylated glycans) significantly reduced the number of skeletal tumours formed (Mann–Whitney test, p = 0.0454), the incidence of tumour in left tibias (Chi-square test, p = 0.0455), and significantly increased survival time in mice (Log-rank test, p = 0.012). ( i ) Micro-CT analysis demonstrated that P-SiaFNEtoc significantly alleviated bone destruction in the trabecular bone of tibias and increased trabecular bone volume (BV/TV, p = 0.0211) and trabecular number (Tb. N, p = 0.035) (n = 9, unpaired t test, ∗p < 0.05). Representative images are shown. Scale bar is 200 μm.

Article Snippet: Slides were incubated for 3 h at room temperature with FITC-conjugated SNA lectin (Vector labs, FL-1301-2) at 1:500 or FITC-conjugated MAL I Lectin (Vector labs, FL-1311-2) at 1:500.

Techniques: Inhibition, Flow Cytometry, Control, Binding Assay, Luciferase, Injection, In Vivo, Imaging, MANN-WHITNEY, Micro-CT