Journal: PLoS ONE

Article Title: Electroacupuncture Ameliorates Acute Renal Injury in Lipopolysaccharide-Stimulated Rabbits via Induction of HO-1 through the PI3K/Akt/Nrf2 Pathways

doi: 10.1371/journal.pone.0141622

Figure Lengend Snippet: β-actin was monitored as the internal standard to ensure similar gel loading of the starting materials in each sample. Blot images were cropped for comparison (E). Electroacupuncture treatment significantly increased the levels of phospho-Akt protein, HO-1 protein, Nrf2 total and nucleoprotein compared with group L or group SEL. In wortmannin-treated rabbits, phosphorylation of Akt was significantly decreased while HO-1 protein, Nrf2 total and nucleoprotein was slightly decreased compared with group EL. All values were expressed as mean ±SD (n = 10; *P<0 . 05; **P<0 . 01 , using one-way ANOVA and the Bonferroni test for multiple comparisons). The blots were representative of three independent experiments.

Article Snippet: After protein blockade, the sections were incubated overnight at 4°Cwith the polyclonal Nrf2 primary antibody conjugated to FITC (dilution 1:150, Biorbyt, UK).

Techniques: Comparison, Phospho-proteomics

Journal: PLoS ONE

Article Title: Electroacupuncture Ameliorates Acute Renal Injury in Lipopolysaccharide-Stimulated Rabbits via Induction of HO-1 through the PI3K/Akt/Nrf2 Pathways

doi: 10.1371/journal.pone.0141622

Figure Lengend Snippet: (original magnification×400): (A) The pictures of immunofluorescence staining. Green stands for Nrf2-FITC stained sections, while blue stands for images of DAPI stained nuclei. The overlay color of blue staining in nucleus, accompanied with green staining both in cytoplasm and nucleus was considered to be positive. (B) Quantification of nuclear localization of Nrf2 protein among six groups. Electroacupuncture protocols dramatically increased translocation of Nrf2 from cytoplasm into nucleus compared with group L or SEL. To some degree, pretreatment with wortmannin counteracted nuclear accumulation of Nrf2 protein, while wortmannin alone had no effects. Data were representative of three independent experiments. Values were expressed as mean ± SD (n = 10; **P<0 . 01 , using one-way ANOVA and the Bonferroni test for multiple comparisons).

Article Snippet: After protein blockade, the sections were incubated overnight at 4°Cwith the polyclonal Nrf2 primary antibody conjugated to FITC (dilution 1:150, Biorbyt, UK).

Techniques: Immunofluorescence, Staining, Translocation Assay

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