Journal: Molecular Neurodegeneration

Article Title: Immunochemical characterization on pathological oligomers of mutant Cu/Zn-superoxide dismutase in amyotrophic lateral sclerosis

doi: 10.1186/s13024-016-0145-9

Figure Lengend Snippet: Anti-SOD1 int antibody exclusively recognizes soluble disulfide-crosslinked SOD1 oligomers in vitro. a The antibodies were tested for their specific reactivities to soluble disulfide-crosslinked oligomers (black filled bars) over Cu,Zn-SOD1(WT) S-S (open bars) and E,E-SOD1(A4V) S-S (gray filled bars) by indirect ELISA. Antisera were either affinity-purified with the corresponding peptides (w/o absorption) or first absorbed with SOD1(WT) S-S and then affinity-purified with the peptides (w/ absorption). Anti-SOD1 48–53 antibody obtained after the absorption exclusively reacted with soluble disulfide-crosslinked oligomers and called anti-SOD1 int antibody. b - d The reactivities of b anti-SOD1 int , c USOD-like, and d SEDI-like antibody were examined with indirect ELISA. Several forms of SOD1 (WT, A4V, G37R, G85R) with a distinct metallation/disulfide status, soluble disulfide-crosslinked oligomers and insoluble amyloid-like aggregates were prepared and fixed on an ELISA plate. The ELISA signal was represented as a ratio against that obtained using BSA. Three independent experiments were performed to estimate error bars (standard deviation). Fixation of equal amounts of SOD1 proteins on each well of an ELISA plate was confirmed by ELISA using polyclonal anti-SOD1 antibody (FL-154, Santa Cruz Biotechnology), which is shown in Additional file : Figure S5

Article Snippet: After six washes with TBS-T, either antibody purified in this study, polyclonal anti-human SOD1 (FL-154, Santa Cruz Biotechnology), USOD (#SPC-205, StressMarq Bioscience), or SEDI (#SPC-206, StressMarq Bioscience) antibody was added as a primary antibody (0.2 μg/mL) and incubated for an hour at room temperature, which was then followed by secondary antibody with horseradish peroxidase (goat anti-rabbit IgG, 1:1,000; Thermo Scientific) for an hour at room temperature.

Techniques: In Vitro, Indirect ELISA, Affinity Purification, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Acta biochimica et biophysica Sinica

Article Title: Vaccarin suppresses diabetic nephropathy through inhibiting the EGFR/ERK1/2 signaling pathway.

doi: 10.3724/abbs.2024141

Figure Lengend Snippet: Figure 3. VAC alleviated the renal inflammatory response and oxidative stress in T2DM mice (A) MDA contents in diabetic kidney tissues. n=6. (B) GSH-Px activity in diabetic kidney tissues. n=6. (C) Averaged fluorescence intensity of DHE fluorescence in diabetic kidney tissues. n=6. (D) Averaged fluorescence intensity of DCFH-DA fluorescence staining of diabetic kidney tissues. n=6. (E) DHE fluorescence staining or DCFH-DA fluorescence staining of diabetic kidney tissues. Scale bar: 100 μm. (F) F4/80 staining of diabetic kidney tissues. Scale bar: 50 μm, and the average fluorescence intensity of F4/80-expressing diabetic kidney tissues is shown. (G‒M) Representative blot images and quantitative analysis of phosphorylated NFκB P65, Nrf2, catalase, SOD3, SOD2 and SOD1. n=4. *P<0.05, **P<0.01, ***P<0.001 vs Ctrl. #P<0.05, ##P<0.01, ###P<0.001 vs DN.

Article Snippet: Primary antibodies against catalase, SOD1, SOD2 and SOD3 were procured from Boster Biological Technology (Wuhan, China).

Techniques: Activity Assay, Fluorescence, Staining, Expressing

Journal: Acta biochimica et biophysica Sinica

Article Title: Vaccarin suppresses diabetic nephropathy through inhibiting the EGFR/ERK1/2 signaling pathway.

doi: 10.3724/abbs.2024141

Figure Lengend Snippet: Figure 5. VAC alleviated the inflammatory response and oxidative stress in diabetic kidneys HK-2 cells were preincubated with 5 μM VAC for 12 h, followed by exposure to 35 mM HG for 48 h. (A–C) Relative mRNA levels of IL-1β, VCAM-1 and COX2. (D-F) DHE fluorescence staining or DCFH-DA fluorescence staining of HK-2 cells. Scale bar: 100 μm. (G–K) Relative mRNA levels of Nrf2, catalase, SOD3, SOD2 and SOD1. (L–R) Representative blot images and quantitative analysis of phosphorylated NFκB P65, Nrf2, catalase, SOD3, SOD2 and SOD1. *P<0.05, **P<0.01, ***P<0.001 vs NG. #P<0.05, ##P<0.01, ###P<0.001 vs HG. n=4.

Article Snippet: Primary antibodies against catalase, SOD1, SOD2 and SOD3 were procured from Boster Biological Technology (Wuhan, China).

Techniques: Fluorescence, Staining

Journal: Acta biochimica et biophysica Sinica

Article Title: Vaccarin suppresses diabetic nephropathy through inhibiting the EGFR/ERK1/2 signaling pathway.

doi: 10.3724/abbs.2024141

Figure Lengend Snippet: Figure 8. VAC ameliorated fibrosis, the inflammatory response and oxidative stress in HG-induced HK-2 cells exposed to the EGFR inhibitor AG1478 or the ERK inhibitor U0126 (A–D) Relative mRNA levels of collagen-1, TGF-β1, α-SMA and E-cadherin in HK-2 cells. (E–G) Relative mRNA levels of IL-1β, VCAM-1 and COX-2 in HK-2 cells. (H) Averaged fluorescence intensity of DHE fluorescence in HK-2 cells. (I) DHE staining was performed on HG-induced HK-2 cells. Scale bar: 100 μm. (J–P) Representative blot images and quantitative analysis of phosphorylated and total NFκB P65, Nrf2, catalase, SOD3, SOD2 and SOD1. *P<0.05, **P<0.01, ***P<0.001 vs NG. #P<0.05, ##P<0.01, ###P<0.001 vs HG. n=4.

Article Snippet: Primary antibodies against catalase, SOD1, SOD2 and SOD3 were procured from Boster Biological Technology (Wuhan, China).

Techniques: Fluorescence, Staining

Journal: Redox Biology

Article Title: The loss of pancreatic islet NADPH oxidase (NOX)2 improves islet transplantation

doi: 10.1016/j.redox.2022.102419

Figure Lengend Snippet: The loss of NOX2 increases the expression of insulin and antioxidative proteins in hypoxic islets. ( A ) Representative Western blot analysis of (from top to bottom) Nrf2, β-actin, HO-1, SOD2, SOD1 and insulin from whole cell extracts of isolated hypoxic WT and Nox2 −/− islets. ( B ) Quantitative analysis of insulin expression (Fold change) (n = 5 each). Mean ± SEM. *P < 0.05 vs. WT. ( C ) Quantitative analysis of SOD1 expression (Fold change) (n = 3 each). Mean ± SEM. *P < 0.05 vs. WT. ( D ) Quantitative analysis of SOD2 expression (Fold change) (n = 3 each). Mean ± SEM. *P < 0.05 vs. WT. ( E ) Quantitative analysis of HO-1 expression (Fold change) (n = 5 each). Mean ± SEM. *P < 0.05 vs. WT. ( F ) Quantitative analysis of Nrf2 expression (Fold change) (n = 5 each). Mean ± SEM. *P < 0.05 vs. WT.

Article Snippet: The anti-SOD1 (TA321133) and anti-SOD2 (TA321189) antibodies were purchased from OriGene (Rockville, USA).

Techniques: Expressing, Western Blot, Isolation

Journal: Acta Neuropathologica Communications

Article Title: Quantitative patterns of motor cortex proteinopathy across ALS genotypes

doi: 10.1186/s40478-020-00961-2

Figure Lengend Snippet: Summary of all cases used in this study. See supplementary data for subgroup analyses

Article Snippet: SOD1 d , StressMarq Bio , SPC-206 , Rabbit , Polyclonal , 1:1000 , –.

Techniques:

Journal: Acta Neuropathologica Communications

Article Title: Quantitative patterns of motor cortex proteinopathy across ALS genotypes

doi: 10.1186/s40478-020-00961-2

Figure Lengend Snippet: Primary antibody clones used in this study

Article Snippet: SOD1 d , StressMarq Bio , SPC-206 , Rabbit , Polyclonal , 1:1000 , –.

Techniques: Clone Assay

Journal: Acta Neuropathologica Communications

Article Title: Quantitative patterns of motor cortex proteinopathy across ALS genotypes

doi: 10.1186/s40478-020-00961-2

Figure Lengend Snippet: Primary motor cortex and spinal cord single-IHC results

Article Snippet: SOD1 d , StressMarq Bio , SPC-206 , Rabbit , Polyclonal , 1:1000 , –.

Techniques:

Journal: Acta Neuropathologica Communications

Article Title: Quantitative patterns of motor cortex proteinopathy across ALS genotypes

doi: 10.1186/s40478-020-00961-2

Figure Lengend Snippet: Pathology of the ALS primary motor cortex is variable both within and across the genotypic spectrum of disease. Relatively little pTDP-43 aggregation was found in a single TARDBP mutation case ( a ), but this was severe in an OPTN mutation case (d; see also supp. Figure ). Insets highlight variance of pTDP-43 morphology between genotypes. Average highest pTDP-43 deposition was seen in sporadic cases, which was statistically higher than in C9-ALS ( g ). Quantification of p62 ( b , e , h ). Levels of p62 correlated with pTDP-43 in sporadic cases but less so in C9ORF72 disease, reflective of the existence of p62-positive dipeptide repeat protein species unique to C9-ALS ( h and j ). Cortical microglial activation was highly variable between genotypes ( i ), and in some cases there was evidence of severe nodular neuronophagia surrounding layer V neurons ( f ). Grey matter CD68 correlated with the extent of pTDP-43 deposition ( k ) in both sporadic and C9ORF72 cases, but this relationship was not recapitulated using p62 and CD68 in SOD1 / FUS cases ( l ). Arrows highlight pathology, asterisks highlight Betz cells. r correlations = Pearson ( j ) or Spearman ( k , l ), results as on figure. Bars in ( i ) represent means and SEM. Best-fit lines are manually added for illustrative purposes. All scale bars = 50 μm

Article Snippet: SOD1 d , StressMarq Bio , SPC-206 , Rabbit , Polyclonal , 1:1000 , –.

Techniques: Mutagenesis, Activation Assay

Journal: Acta Neuropathologica Communications

Article Title: Quantitative patterns of motor cortex proteinopathy across ALS genotypes

doi: 10.1186/s40478-020-00961-2

Figure Lengend Snippet: Spatial and morphological distribution of proteinopathy across genotypes in the ALS primary motor cortex. Sporadic ALS exhibits NCI and oligodendroglial pathology across layers I-VI as well as the subcortical white matter ( a - d ). The distribution pattern of pTDP-43 pathology was not entirely dissimilar between sporadic and C9-ALS ( e - f ), although there was a slight preponderance towards oligodendrocyte inclusions in C9 cases (Fig. g and h). FUS mutation cases demonstrated infrequent, compact p62-positive NCI with occasional granular inclusions, as well as occasional oligodendroglial pathology (i-l). SOD1 mutation cases, by contrast, exhibited granular p62 staining confined to the middle cortical layers with infrequent NCI in layer V, without obvious glial pathology. Arrows highlight respective proteinopathy. Scale bar applicable to all panels = 50 μm

Article Snippet: SOD1 d , StressMarq Bio , SPC-206 , Rabbit , Polyclonal , 1:1000 , –.

Techniques: Mutagenesis, Staining