cwith primary antibodies against ets 1  (Cell Signaling Technology Inc)

Journal: Nano Select

Article Title: Exosome‐Loaded Contact Lenses: A Novel Approach for Sustained Ocular Drug Delivery

doi: 10.1002/nano.70045

Figure Lengend Snippet: FIGURE 4 Material characterization: (a) Water content assessment. (b) Optical properties assessment. (c) Wettability assessment. (d) Laser intensity assessment. (e) Detection of GAPDH in exosomal RNA at different lens incubation time points (after each timepoint, the supernatant was collected, and unbound exosomes were pelleted by centrifugation. RNA was extracted using TRIzol Reagent, converted to cDNA, and analyzed by qPCR using 10 ng of input cDNA and GAPDH-specific primers. The graph shows the mean Ct values (±standard deviation) for each condition based on four replicates). (f) Western blot detection of the exosomal marker CD63 (Exosomes were lysed and subjected to SDS-PAGE, followed by transfer to a nitrocellulose membrane. The membrane was probed with CD63 [E1W3T] Rabbit mAb [#52090, Cell Signaling Technology], and the band was visualized using HRP-conjugated antirabbit IgG and chemiluminescent detection on a Bio-Rad imaging system. A strong band corresponding to CD63 was observed.).

Article Snippet: Membranes were blocked with 5% nonfat drymilk in TBST for 1 h at room temperature, then incubated overnight at 4◦Cwith CD63 (E1W3T) Rabbit mAb (1:1000; Cell Signaling Technology, #52090), a commonly used exosomal marker, diluted in 5% milk in TBST.

Techniques: Incubation, Centrifugation, Standard Deviation, Western Blot, Marker, SDS Page, Membrane, Imaging

Journal: Alzheimer's & dementia : the journal of the Alzheimer's Association

Article Title: Pathological angiogenesis was associated with cerebrovascular lesion and neurodegeneration in Alzheimer's disease.

doi: 10.1002/alz.14521

Figure Lengend Snippet: FIGURE 1 Cerebral vessels lesion and elevated CD31 levels related to tau pathology in human AD brains. Representative western blots of the level of CD31 (endothelial marker), αSMA (smooth muscle marker), and PDGFRβ (pericyte marker) in separated cerebral vessels of human non-ADD (n = 9) and AD brains (n = 20). Relative expression of αSMA and PDGFRβ normalized to CD31. (A) While both αSMA/CD31 and PDGFRβ/CD31 in cerebral vessels were reduced in human AD brains compared to non-AD brains, CD31 levels showed no difference between the two groups. (B) Schematic of extraction of soluble (TBS fraction), membrane (TBSX fraction) and insoluble proteins (GuHCl fraction) from frozen human brains by a three–step ultracentrifugation. (C) Quantification of occludin, Claudin-5, and CD31 in TBSX fraction by ELISA. Both occludin/CD31 and Claudin-5/CD31 were reduced in human AD brains (n = 7) compared to non-ADD brains (n = 24). (D) Quantification of Aβ42, Aβ40 and pTau181 in GuHCl fraction by MSD. The high pathological group is defined as having levels of Aβ42, Aβ40, or pTau181 greater than the median of the AD group. (E) Univariate or multivariate analyses was used to test associations between Aβ42, Aβ40, pTau181 in GuHCl fraction and CD31 in TBSX fraction with age, sex, and APOE genotype as covariates. Group differences were assessed using ANCOVA adjusted age, sex, and APOE genotype, following by Bonferroni multiple comparison tests. *p < 0.05, **p < 0.01. AD, Alzheimer’s disease; ANCOVA, analysis of covariance; APOE, apolipoprotein E.

Article Snippet: The sections were then blocked with blocking buffer (5% normal goat serum, 0.3% Triton ×-100 in phosphate buffered saline [PBS]) for 1 h at room temperature and incubated overnight at 4◦Cwith primary antibodies against CD31 (CST, 3528S, 1:200) and CD13 (Proteintech, 66211-1-Ig, 1:200).

Techniques: Western Blot, Marker, Expressing, Extraction, Membrane, Enzyme-linked Immunosorbent Assay, Comparison