Journal: International Journal of Nanomedicine

Article Title: Glycosylation-Engineered Platelet Membrane-Coated Interleukin 10 Nanoparticles for Targeted Inhibition of Vascular Restenosis

doi: 10.2147/IJN.S423186

Figure Lengend Snippet: Schematic diagram and identification of glycosylation engineering of platelet membrane glycoproteins and the characterization of GE-PNP. ( A ) Schematic diagram of glycosylation engineering. ( B and C ) Flow cytometry detected lectins binding form in neuraminidase group (NEU) and neuraminidase + fucose engineered(FE) group. ( D ) Transmission electron microscopy of empty PLGA nanoparticles (NPs), IL10-NPs, IL10-PNPs, and IL10-GE-PNPs. Scale bar = 100μm. ( E ) Coomassie stain to visualize platelet vesicles and empty PNPs/GE-PNPs on SDS-PAGE at equal protein levels. ( F ) Western blot to identify platelet membrane glycoprotein in platelet vesicles and PNPs/GE-PNPs. ( G ) Size and ( H ) Zeta potential of the empty PLGA core, IL10-NP, empty PNP, IL10-PNP, empty GE-PNP, and IL10-GE-PNP are evaluated. ( I ) Monitoring the diameter of GE-PNP in deionized water on day 0 and day 7. ( J ) Determining the encapsulation efficiency and ( K ) loading capacity of IL10 in IL10-NP, IL10-PNP, and IL10-GE-PNP. ( L ) Measuring the in vitro cumulative release of IL10-NP, IL10-PNP, and IL10-GE-PNP after incubation at 37°C in PBS Data are presented as mean ± SD, ***p < 0.001, n=3.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated lectins (Vector Laboratories, USA) were used to bind to specific sugars, including Ricinus communis Agglutinin I (RCA I), which binds to β-galactose to identify the removal of sialic acids; Ulex europaeus agglutinin-I (UEA-I), which binds to α (1,2)-fucose; and Lotus tetragonolobus lectin (LTL), which binds to α (1,3)-fucose., Platelets were incubated with FITC-conjugated lectins for 20 min at room temperature.

Techniques: Membrane, Flow Cytometry, Binding Assay, Transmission Assay, Electron Microscopy, Staining, SDS Page, Western Blot, Encapsulation, In Vitro, Incubation

Journal: Acta Neuropathologica Communications

Article Title: A single mild juvenile TBI in male mice leads to regional brain tissue abnormalities at 12 months of age that correlate with cognitive impairment at the middle age

doi: 10.1186/s40478-023-01515-y

Figure Lengend Snippet: Vascular change was observed in the SI/NB 12 m after jmTBI. A Blood vessels were stained with tomato-lectin in both groups. A decrease in vessel diameter was observed in the SI/NB of the jmTBI ( A2 , A4 ) compared to sham group ( A1 , A3 ). B Significant increase of the proportion of small blood vessel with diameter between [0–5μm] in jmTBI compared to sham mice in both ipsilateral and contralateral sides of the SI/NB

Article Snippet: Fluorescein-conjugated tomato lectin (1:500, Vector, Vector Laboratories, Burlingame, CA) was used to detect vessels.

Techniques: Staining

Journal: mBio

Article Title: Genetic Ablation of a Female-Specific Apetala 2 Transcription Factor Blocks Oocyst Shedding in Cryptosporidium parvum

doi: 10.1128/mbio.03261-22

Figure Lengend Snippet: C. parvum cgd4-1110 encodes an AP2 DNA binding protein that is expressed exclusively in female gametes. (A) The locus of cgd4_1110 was modified to append a 3xHA epitope tag to the coding sequence. The resulting parasite strain was used to infect HCT-8 cell cultures and analyzed by immunofluorescence assays (IFA) at 24 and 48 h postinfection. The transgene was detected with antibodies to HA (red), and all parasites were labeled with Vicia villosa lectin (VVL, green). (B) Quantification of HA staining as a proportion of all parasites. (C) IFA of cultures infected with cgd4_1110-mNG showing asexual meronts, a female gamete, and a male gamont at 48 h. mNeon green (mNG) fluorescent protein is shown in green, DNA stained with DAPI in blue, and TrpB (labeling all parasites) in red. The single nucleus of the female gamete is highlighted with an arrowhead. (D) cgd4_1110-3xHA (red) expressing parasites stained for the female marker DMC1 (note that all HA-positive parasites are also DMC1 positive).

Article Snippet: Parasites were counterstained with anti-HA antibody and fluorescein-conjugated Vicia villosa lectin (Vector Laboratories).

Techniques: Binding Assay, Modification, Sequencing, Immunofluorescence, Labeling, Staining, Infection, Expressing, Marker