Journal: Frontiers in Endocrinology

Article Title: Integration of Metabolomics and Proteomics in Exploring the Endothelial Dysfunction Mechanism Induced by Serum Exosomes From Diabetic Retinopathy and Diabetic Nephropathy Patients

doi: 10.3389/fendo.2022.830466

Figure Lengend Snippet: Serum exosomes from diabetic retinopathy and nephropathy patients could induce endothelial dysfunction. (A) The typical pathological kidney images and fundus images obtained from healthy people and a diabetic microvascular disease patient. PAS : The DMC biopsy sample showed proliferation and swelling of the endothelial cell (blue arrow), reduplication (double contour appearance) of the glomerular basement membrane (red arrow), mesangial expansion (black arrow), arteriolar hyalinosis (green arrow). Scale bar: 50 μm. Methenamine silver: Renal biopsy samples from the DMC patient show the swelling of the endothelial cells (red arrow) and reduplication (double contour appearance) of the glomerular basement membrane (black arrow). Scale bar: 50 μm. Electron microscopy: The slice from the DMC patient showed mesangial expansion (red arrow). Scale bar: 5 μm. Fundus: The fundus from DMC showed microhemangioma (green arrow), retinal exudates (red arrow), intraretinal hemorrhage (black arrow), and intraretinal microvascular abnormalities (IRMAs; violet arrow). (B) Identification of serum exosomes from CON patients and DRDN patients by transmission electron microscopy (TEM). Scale bar: 200 nm. (C) Western blotting was used to look for the exosomal markers CD9, CD63, and TSG101 in exosome samples. (D) Analysis of the size distribution of exosomes from patients using the NanoSight technology; the average size of serum exosomes was 107.5 ± 55.2 nm. (E) Exosome tracing experiment captured by confocal microscope. Blue for DNA dyed by DAPI and green for exosomes derived from DR+DN patients dyed by PKH67. HGECs were subjected to 6, 12, and 24 h of incubation with exosomes. Scale bar: 25 μm. PAS, Periodic acid–Schiff stain; DMC, diabetic microvascular complications; CON, healthy controls; DRDN, patients diagnosed with both diabetic retinopathy and diabetic nephropathy.

Article Snippet: The primary antibodies were anti-CD9 (Abcam, Hong Kong, China; # ab92726), anti-CD63 (Abcam, ab216130), anti-TSG101 (Sino Biological, Beijing, China; #102286-T38), anti-CD31 (Zen-Bioscience, Chengdu, China; #383815), anti-von Willebrand factor (vWF), (Abcam, #ab154193), anti-intercellular adhesion molecule 1 (ICAM-1) (Abcam, #ab171123), anti-vascular cell adhesion protein 1 (VCAM-1) (Abcam, #ab134047), and anti-Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) (Goodhere, Hangzhou, China; #AB-P-R 001).

Techniques: Membrane, Electron Microscopy, Transmission Assay, Western Blot, Microscopy, Derivative Assay, Incubation, Staining

Journal: Bioengineered

Article Title: Tumor suppressing role of serum-derived exosomal microRNA-15a in osteosarcoma cells through the GATA binding protein 2/murine double minute 2 axis and the p53 signaling pathway

doi: 10.1080/21655979.2021.1987092

Figure Lengend Snippet: miR-15a from serum-derived exosomes can be internalized by OS cells. a, existence of miR-15a in exosomes predicted using the EVmiRNA system; b, diameter of the extracted particles evaluated by NTA; c, protein levels of TSG101 and CD81 in the particles determined by RT-qPCR; d, morphology of the particles observed under a TEM; e, miR-15a in extracted exosomes examined by RT-qPCR; f, internalization of the exosomes by OS cells confirmed by fluorescence tracking; g, miR-15a expression in OS cells after serum-derived exosome treatment determined by RT-qPCR (* p < 0.05, two-way ANOVA)

Article Snippet: The membranes were blocked with 5% (v/v) skimmed milk for 1 h and incubated with primary antibodies at 4°C for 16 h. The antibodies used were Bcl-2 (ab692, 1:500, Abcam Inc., Cambridge, MA, USA), Bax (ab32503, 1:2,000, Abcam), GAPDH [(#3683, 1:1,000, Cell Signaling Technology (CST), Beverly, MA, USA)], cyclin E1 (ab33911, 1:2,000, Abcam), cyclin-dependent kinase 2 (CDK-2, ab32147, 1:1,000, Abcam), CDK-4 (ab199728, 1:2,000, Abcam), cyclin D1 (ab226977, 1:2,000, Abcam), p53 (sc-126, 1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA), tumor susceptibility gene 101 (TSG101, sc-7964, 1:1,000, Santa Cruz Biotechnology), and CD81 (sc-23,962, 1:500, Santa Cruz Biotechnology).

Techniques: Derivative Assay, Quantitative RT-PCR, Fluorescence, Expressing

Journal: Separations

Article Title: Alternative Method for HDL and Exosome Isolation with Small Serum Volumes and Their Characterizations

doi: 10.3390/separations8110204

Figure Lengend Snippet: Figure 6. The protein pattern and biomarker of the exosomes isolated from different sample volumes. A protein ladder reference (Vivantis Technologies, Shah Alam, Selangor Darul Ehsan, Malaysia, cat no. PR0623) had 2 reference bands (25 and 72 kDa) coupled with blue chromophore for easy identification (A). SDS-PAGE stained with Coomassie blue G250 showed multiple bands with the same patterns (B). The Western blot results showed a single band of TSG101 at about 48.9 kDa exhibiting the exosome biomarker (C). Thirty micrograms of protein were loaded for both SDS-PAGE and the Western blot. All experiments above were run with a reducing agent. M = Marker of protein molecular weight.

Article Snippet: The primary antibodies, human ApoA-1 antibody (cat no. MAB 36641-SP) and human TSG101 antibody (cat no. orb337267), were supplied by R&D System, Inc., Minneapolis, MN, USA and Biorbyt, Cambridge, Cambridgeshire, UK, respectively.

Techniques: Biomarker Discovery, Isolation, SDS Page, Staining, Western Blot, Marker, Molecular Weight

Journal: Advances in Dermatology and Allergology/Postȩpy Dermatologii i Alergologii

Article Title: Modulation of dermal equivalent of hypothalamus-pituitary-adrenal axis in mastocytosis

doi: 10.5114/ada.2021.107933

Figure Lengend Snippet: List of antibodies used in the study

Article Snippet: Anti-MCIR (Rb) , Abcam , ab125031 , 1 : 100 , ImmPRESS™ HRPAnti-Rabbit IgG (Peroxidase) Polymer Detection Kit.

Techniques: