ac 4 mannaz  (MedChemExpress)

Journal: EJNMMI Radiopharmacy and Chemistry

Article Title: Aluminum-[ 18 F]fluoride radiolabeling of triarylphosphines for cell labeling via the perfluoroaryl azide Staudinger ligation

doi: 10.1186/s41181-025-00409-9

Figure Lengend Snippet: A . In vitro cytotoxicity of compound 1 in human Jurkat T lymphoblasts after 24, 48, and 72 h incubation at 10–250 μM. Negative (–) and positive ( +) controls were complete medium with 0.5% v/v DMSO and lysis Buffer (1:10 v/v), respectively. Columns represent mean ± s.d. (n = 4). The statistical significance was analyzed using unpaired Student’s t -test versus the negative control (* p < 0.05, ** p < 0.01, *** p < 0.001, and n.s. = not significant). The dashed pink line indicates 80% cell viability, the threshold for acceptable cytocompatibility. B – D . Flow cytometry analysis of live Jurkat cells after a 72 h treatment with 1 × PBS (dotted line in B and dashed line in C ) or MGE with Ac 4 MAnNAz (solid line in B – D ) or 1 (complex line in D ), followed by incubation with 1 × PBS (dotted line in B ) or 1 µM phosphine-PEG3-biotin (solid line in B – D and complex line in D ) and finally incubation with streptavidin-FITC (all lines)

Article Snippet: Jurkat cells (5 × 10 5 cells in 1.5-mL complete medium per well) were seeded in a non-treated flat-bottom polystyrene 6-well plate (Avantor, Radnor, PA, USA) and incubated in triplicates (n = 3) with 50 μM of 1 , 50 μM of Ac 4 ManNAz (Lumiprobe, Westminster, MD, USA) or 0.5% (v/v) DMSO in complete medium for 72 h in a cell incubator set at 37 oC with 5% CO 2 and 95% relative humidity.

Techniques: In Vitro, Incubation, Lysis, Negative Control, Flow Cytometry

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: GlycoRNA-L and glycoRNA-S mediate human monocyte adhesion via binding to Siglec-5

doi: 10.1016/j.bbamcr.2025.120017

Figure Lengend Snippet: Expression of glycoRNA-L and glycoRNA-S in human monocytes. (A) THP1 cells were treated with or without Ac 4 ManNAz. RNAs were extracted from the cells and treated with or without RNase. Then the RNA samples reacted with DBCO-PEG4-biotin. RNAs were analyzed on an agarose gel (left) and then the blot was probed with an anti-biotin antibody (right). Representative images are shown. (B) 10 μg of biotinylated total RNAs from THP1 cells were incubated with 1 μl of RNase cocktail (containing RNase A 0.5 U/μ/ and RNase T 20 U/μl), DNase I (1 U/μl), PNGase F (1 U/μl) or Proteinase K (2 μg/μl) at 37 °C for 10 min. After cleaning up by zymo column, the RNA samples were analyzed by northern blot as above. (C) Blotting of cellular or in vitro Ac 4 ManNAz-labeled RNA. Cells were treated with 100 μM Ac 4 ManNAz for 24 h while native RNA in vitro was treated with 0, 5, 10 and 20 mM of Ac4ManNAz at 37 °C for 2 h. (D) THP1 cells were treated with Ac 4 ManNAz and different doses of NIG-1 or kifunensine as indicated. RNAs were extracted from the cells and analyzed by northern blot. Representative images are shown. (E) RNA from different cells treated with 100 μM Ac 4 ManNAz for 24 h and then analyzed by the same procedure as above. Representative images are shown. (F) RNA from THP1 or hPBMC (ATCC) treated with 100 μM Ac 4 ManNAz plus or not plus LPS (1 μg/ml) for 24 h and then analyzed by the same procedure as above. Representative images are shown.

Article Snippet: Stocks of N -azidoacetylmannosamine-tetraacylated (Ac 4 ManNAz or ManNAz, Tocris Bioscience, Cat#:7479) were made to 500 mM in sterile dimethyl sulfoxide (DMSO).

Techniques: Expressing, Agarose Gel Electrophoresis, Incubation, Northern Blot, In Vitro, Labeling

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: GlycoRNA-L and glycoRNA-S mediate human monocyte adhesion via binding to Siglec-5

doi: 10.1016/j.bbamcr.2025.120017

Figure Lengend Snippet: Characterization of structures of glycoRNA-L and glycoRNA-S in THP1 cells. (A) THP1 cells were labeled by 100 μM of Ac 4 ManNAz (ManNAz), GalNAz, GlcNAz and FucAz for 24 h respectively. RNAs were extracted from the cells. Then the RNA samples reacted with DBCO-PEG4-biotin. RNAs were analyzed on an agarose gel (left) and then blotted with an anti-biotin antibody (right). Representative images are shown. (B) Blot of RNA from HeLa cells labeled by same reagents as above. (C) THP1 cells were pretreated with or without NIG-1 (10 μM/L) or kifunensine (5 μM/L) for 1 h and then incubated with GalNAz (100 μM/L) for 24 h. The RNA was isolated and analyzed as described above. (D) 10 μg biotinylated total RNA from Ac 4 ManNAz-labeled THP1 cells were treated with 1 μl RNase cocktail and then directly loaded on the denaturing gel and analyzed by same procedure as above.

Article Snippet: Stocks of N -azidoacetylmannosamine-tetraacylated (Ac 4 ManNAz or ManNAz, Tocris Bioscience, Cat#:7479) were made to 500 mM in sterile dimethyl sulfoxide (DMSO).

Techniques: Labeling, Agarose Gel Electrophoresis, Incubation, Isolation