Journal: Nature Communications

Article Title: Mammalian brain glycoproteins exhibit diminished glycan complexity compared to other tissues

doi: 10.1038/s41467-021-27781-9

Figure Lengend Snippet: Protein lysate from a representative male mouse cortex and cerebellum with human plasma as a positive control was treated with or without PNGase F and visualized using biotinylated lectins (ConA, GNL, PHA-E, AAL, RCA, and SNA) in addition to immunoblotting for actin and staining for total protein. Non-specific binding of lectins to PNGase F is noted by an asterisk (*) near 35 kDa, as shown in the Total Protein stain. Protein blotting of brain lysate with each lectin has been repeated at least three times each with similar results. A schematic with common lectin-binding sites is shown for reference. Source data are provided as a Source Data file.

Article Snippet: Human Brain Cerebral Cortex Whole Tissue Lysate was purchased from Novus Biologicals (#NB820-59182), with 1mg used for glycomic analysis as described below.

Techniques: Clinical Proteomics, Positive Control, Western Blot, Staining, Binding Assay

Journal: Scientific Reports

Article Title: Multicellular Self-Assembled Spheroidal Model of the Blood Brain Barrier

doi: 10.1038/srep01500

Figure Lengend Snippet: (a–c) Flow cytometry analysis of confluent (a) hpBECs (green), (b) hpPs (red) and (c) hpAs (blue). Staining was done with indicated antibodies or relevant isotypes (grey population, grey histogram). One representative experiment of three is shown. (d) Examination of hpBEC tube formation (green), hpPs (red) and hpAs (blue) tube association on matrigel matrix using different fluorescent markers′, 18 hours after seeding. The images were taken as an overlay of five 1 um thick confocal sections. Single channels (D1-3) and the merged image of all colors (D4) are displayed. White arrows in the upper right red channel image (D2) indicate hpPs aligned along an hpBECs tube and the white arrows in the lower right merged image (D4) point out the loosely associated hpAs. Bar equals 100 um.

Article Snippet: Human cerebral astrocytes (hpAs) and human brain vascular pericytes (hpPs) were acquired from ScienCell, USA.

Techniques: Flow Cytometry, Staining

Journal: Scientific Reports

Article Title: Multicellular Self-Assembled Spheroidal Model of the Blood Brain Barrier

doi: 10.1038/srep01500

Figure Lengend Snippet: Representative laser confocal images of fluorescently labeled hpBECs (green), hpPs (red) and hpAs (blue) single and co-culture spheroids. Cells were stained using cell tracker dyes for long term tracing of living cells. (a) hpBECs, (b) hpPs single-culture and (c) hpBEC/hpPs co-culture spheroids cultured for 3 days. (d) hpPs/hpAs co-culture spheroids cultured for 1 day prior to addition of (e) hpBECs for additional 2 days. The pictures were taken as an overlay of five 1 um thick confocal sections through the middle of the spheroid. A total of 20 spheroids in three independent experiments were analyzed for each culture condition. Bar equals 50 um. f1-f4 shows a spontaneously formed spheroid where the hpBECs is selectively shown in green (f1), hpPs selectively shown red (f2), hpAs selectively shown in blue (f3) and the complete spheroid (f4) composed of all the three different cell types.

Article Snippet: Human cerebral astrocytes (hpAs) and human brain vascular pericytes (hpPs) were acquired from ScienCell, USA.

Techniques: Labeling, Co-Culture Assay, Staining, Cell Culture

Journal: Scientific Reports

Article Title: Multicellular Self-Assembled Spheroidal Model of the Blood Brain Barrier

doi: 10.1038/srep01500

Figure Lengend Snippet: HpBECs single cultures, hpBECs/hpPs and hpBECs/hpPs/hpAs co-cultures grown as (a & b) spheroids or (c) in the trans-well set-up for 3 days were stained with indicated cell type specific antibodies and analyzed by flow cytometry. (b)Spheroid derived CD31+ & CD140b- gated hpBECs or (c) hpBECs harvested from the trans-well inserts were further examined for the indicated receptor expression. (b & c) Surface receptor expression levels on hpBECs are displayed for single hpBECs culture (black histogram), co-cultured with hpPs (red histogram) or triple co-cultured with hpPs and hpAs (blue histogram). Data are representative of two experiments with 1000 spheroids respectively 3 trans-well filters per individual staining. (MFI = mean fluorescence intensity).

Article Snippet: Human cerebral astrocytes (hpAs) and human brain vascular pericytes (hpPs) were acquired from ScienCell, USA.

Techniques: Staining, Flow Cytometry, Derivative Assay, Expressing, Cell Culture, Fluorescence