Journal: Life Science Alliance

Article Title: C9ORF72 -derived poly-GA DPRs undergo endocytic uptake in iAstrocytes and spread to motor neurons

doi: 10.26508/lsa.202101276

Figure Lengend Snippet: APOJ protein levels were measured in three distinct healthy lines of iAstrocytes’ conditioned medium. (A) Regression analysis with the Four-parameter logistic curve-fit (4-PL) method was used to determine the best-fit standard curve based on twofold serial dilutions of APOJ standards and absorbance at 450 nm. (B) iAstrocytes were either left untreated or treated for 24 h with various recombinant DPRs at 1 µM (poly-GA 34 fibrils, poly-GA 34 oligomers, poly-PA 50 oligomers, and poly-GP 24 oligomers), and then APOJ protein levels in their conditioned medium were measured and calculated using the previously generated 4-PL standard curve. Bar graphs of mean ± SEM. One-way ANOVA with Tukey’s multiple-comparisons test (control group is “untreated”). * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001.

Article Snippet: APOJ protein levels were measured using a human APOJ ELISA kit (#KE00110; ProteinTech) following the manufacturer’s instructions.

Techniques: Recombinant, Generated, Control

Journal: Scientific Reports

Article Title: Clusterin mediates hydroquinone-induced cytotoxic responses in HL-60 differentiated cells

doi: 10.1038/s41598-024-82140-0

Figure Lengend Snippet: CLU protein concentration in HL-60 cells exposed to different concentrations of HQ. CLU protein levels were quantified using ELISA after treatment with HQ at concentrations of 0, 10, 25, and 50 µmol/L ( n = 3 per group). The error bars represent SD, and the mean is shown as horizontal bars. The control group (0 µmol/L HQ) exhibited the highest CLU concentration (19.05 ± 2.32 ng/mL), while HQ-treated groups showed a progressive decrease: 14.45 ± 1.23 ng/mL at 10 µmol/L, 9.43 ± 0.40 ng/mL at 25 µmol/L, and 9.46 ± 0.67 ng/mL at 50 µmol/L. Significant differences were determined by ANOVA followed by Tukey post hoc test (* p < 0.05, ** p < 0.01, *** p < 0.001, ns p > 0.05), ns : not significant.

Article Snippet: The concentration of sCLU proteins was measured using the Human Clusterin ELISA Kit (Boster Biological Technology, China) according to the instructions.

Techniques: Protein Concentration, Enzyme-linked Immunosorbent Assay, Control, Concentration Assay

Journal: Scientific Reports

Article Title: Clusterin mediates hydroquinone-induced cytotoxic responses in HL-60 differentiated cells

doi: 10.1038/s41598-024-82140-0

Figure Lengend Snippet: Concentration of sCLU protein in the cell culture supernatant after exposure to varying HQ concentrations. The concentration of sCLU protein in the supernatant of HL -60 cell cultures was determined using ELISA following treatment with HQ at 0, 10, 25, and 50 µmol/L ( n = 3 per group). The control group (0 µmol/L HQ) exhibited the highest concentration (25.88 ± 4.24 ng/mL), while the 10 µmol/L HQ group showed a slight, non-significant decrease (23.73 ± 2.03 ng/mL, p = 0.696). Significant reductions were observed in the 25 µmol/L (17.45 ± 0.36 ng/mL) and 50 µmol/L (12.96 ± 0.67 ng/mL) groups compared to the control (* p < 0.05, *** p < 0.001, ns p > 0.05), ns : not significant.

Article Snippet: The concentration of sCLU proteins was measured using the Human Clusterin ELISA Kit (Boster Biological Technology, China) according to the instructions.

Techniques: Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Serum Proteomic Profiles Reflect the Stages of Myxomatous Mitral Valve Disease in Dogs

doi: 10.3390/ijms24087142

Figure Lengend Snippet: Heat map of 21 identified and quantified serum proteins with significantly different abundances ( p < 0.05, FDR < 0.05) in dogs diagnosed with different stages (B1 (N = 13), B2 (N = 12), and C (N = 13)) of naturally occurring myxomatous mitral valve disease (MMVD) and the healthy/control group (N = 12). N represents the number of serum samples (biological replicates). A heat map was created using the log2FC values (FC = fold change) calculated for master proteins after post-hoc analysis in R. Gene ontology analysis was performed in the PANTHER classification tool with Canis lupus familiaris as the selected organism. The heat map was created in Microsoft Excel. ADIB = adiponectin B; GSN = actin-depolymerizing factor/gelsolin; ITIH1 = inter-alpha-trypsin inhibitor heavy chain 1; C3 = anaphylatoxin-like domain-containing protein/complement C3; RBP4 = plasma retinol-binding protein; PROS1 = vitamin K-dependent protein S; HP = haptoglobin; CLU = clusterin, SERPINF2 = serpin family F member 2; LRG1 = leucine-rich alpha-2-glycoprotein 1; F12 = coagulation factor XII; CCL14 = C-C motif chemokine; C7 = complement C7; CLEC3B = C-type lectin domain family 3 member B/tetranectin; TFRC = transferrin receptor protein 1; ITIH2 = inter-alpha-trypsin inhibitor heavy chain 2; C4BPA = complement component 4 binding protein alpha; PEPD = peptidase D; IGFBP3 = insulin-like growth factor binding protein 3; APBB1P = amyloid beta precursor protein binding family B member 1; ITIH4 = inter-alpha-trypsin inhibitor heavy chain 4.

Article Snippet: A canine-specific ELISA kit targeting clusterin (CLU, catalogue number: CSB-E13770c) (Cusabio, Houston, TX, USA, https://www.cusabio.com , accessed on 14 March 2022) was purchased.

Techniques: Control, Clinical Proteomics, Binding Assay, Coagulation, Protein Binding

Journal: International Journal of Molecular Sciences

Article Title: Serum Proteomic Profiles Reflect the Stages of Myxomatous Mitral Valve Disease in Dogs

doi: 10.3390/ijms24087142

Figure Lengend Snippet: Analytical validation of proteomics results. Serum concentrations of haptoglobin (HP) measured by haptoglobin spectrophotometric assays ( a ) and clusterin assessed through an ELISA ( b ). Western blotting and example of relative density comparison of serum peptidase D ( c ) and gelsolin ( d ) proteins. N represents the number of serum samples (biological replicates). HP concentrations were significantly higher (Kruskal–Wallis test, p value = 0.0004) in diseased dogs compared to the healthy/control group. Moreover, HP concentrations were significantly lower in dogs with MMVD B2 stage compared to dogs with MMVD stage B1 ( a ). Clusterin concentrations were significantly higher (Kruskal–Wallis test, p value = 0.0082) in dogs diagnosed with MMVD compared to the healthy/control group ( b ). Peptidase D was significantly lower (Kruskal–Wallis test, p value = 0.0007) in diseased dogs compared to the healthy/control group ( c ). Gelsolin was significantly lower (Kruskal–Wallis test, p value = 0.0182) in dogs with MMVD stage B1 compared to the healthy/control group, and although values were lower for dogs with MMVD stage B2, this difference was not significant ( d ). a statistically significantly different from control; b statistically significantly different from the MMVD stage B1; c statistically significantly different from the MMVD stage B2; d statistically significantly different from the MMVD stage C.

Article Snippet: A canine-specific ELISA kit targeting clusterin (CLU, catalogue number: CSB-E13770c) (Cusabio, Houston, TX, USA, https://www.cusabio.com , accessed on 14 March 2022) was purchased.

Techniques: Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Western Blot, Comparison, Control

Journal: Journal of Toxicologic Pathology

Article Title: Comparison of changes in urinary and blood levels of biomarkers associated with proximal tubular injury in rat models

doi: 10.1293/tox.2014-0039

Figure Lengend Snippet: Time course of the appearance of urinary and blood biomarkers in rats given a single administration or 4 administrations of gentamicin (GM, 240 mg/kg/day), rats given a single administration of cisplatin (CDDP, 5 mg/kg) or rats used as the vehicle control. The urinary and serum/plasma levels of Kim-1(a, b), NGAL (c, d), CLU (e, f), CysC (g, h) and β2M (i, j) were assayed at the indicated time points (the day after initial dosing shown as day 2). Individual data and mean values are expressed. The sample numbers were as follows: n = 12 for urinalysis on days 2 and 5 and blood analysis on day 2, except for the CDDP group (n = 10 for urinalysis on day 5), and n = 6 for urinalysis on day 12 and blood analysis on days 5 and 12, except for the CDDP group (n = 4 for urinalysis and blood analysis on day 12). * P< 0.05 and ** P< 0.01 vs. control group (two-tailed Dunnett’s multiple comparison test).

Article Snippet: Measurements of urinary and plasma Kim-1 (R&D Systems, Minneapolis, MN, USA), NGAL (BioPorto Diagnostics, Gentofte, Denmark), CLU (Life Diagnostics, Inc., West Chester, PA, USA) and β2M (Mitsubishi Chemical Medience) were performed using commercial ELISA kits.

Techniques: Two Tailed Test

Journal: Journal of Toxicologic Pathology

Article Title: Comparison of changes in urinary and blood levels of biomarkers associated with proximal tubular injury in rat models

doi: 10.1293/tox.2014-0039

Figure Lengend Snippet: Relationship between the urinary and serum/plasma levels of biomarkers. Pearson’s product-moment correlation coefficient was used to evaluate the linear association between the urinary and blood levels of each biomarker. Kim-1 (GM group, coefficient of correlation (R) = 0.96, P< 0.01; CDDP group, R=0.71, P< 0.01) (a), NGAL (GM group, R=0.89, P< 0.01; CDDP group, R=0.95, P< 0.01) (b), CLU (GM group, R=0.10, P= 0.65; CDDP group, R=0.60, P< 0.01) (c), CysC (GM group, R=0.25, P= 0.26, CDDP group, R=0.40, P= 0.07) (d), β2M (GM group, R=0.33, P= 0.13, CDDP group, R=0.68, P< 0.01) (e).

Article Snippet: Measurements of urinary and plasma Kim-1 (R&D Systems, Minneapolis, MN, USA), NGAL (BioPorto Diagnostics, Gentofte, Denmark), CLU (Life Diagnostics, Inc., West Chester, PA, USA) and β2M (Mitsubishi Chemical Medience) were performed using commercial ELISA kits.

Techniques: Biomarker Assay