Journal: BBA Clinical

Article Title: Novel panel of protein biomarkers to predict response to bortezomib-containing induction regimens in multiple myeloma patients

doi: 10.1016/j.bbacli.2017.05.003

Figure Lengend Snippet: Label-free mass spectrometry data. List of statistically significant discovered proteins using LC-MS analysis. Proteins (in bold) were selected for further validation using ELISAs.

Article Snippet: Four commercially available kits for these four proteins; angiogenin (ANG) [Abcam, UK - ab99970], clusterin (CLU) [R&D system, UK - DCLU00], C-C Motif Chemokine 18 (CCL18) [Abcam, UK - ab100620], and Complement C1q [Abcam, UK - ab170246] were used.

Techniques: Mass Spectrometry, Biomarker Discovery, Variant Assay

Journal: BBA Clinical

Article Title: Novel panel of protein biomarkers to predict response to bortezomib-containing induction regimens in multiple myeloma patients

doi: 10.1016/j.bbacli.2017.05.003

Figure Lengend Snippet: ELISA Data. Mean, SD, Area under the curve (AUC) and p -value for each of the new and standard proteins found in the two groups of patients compared.

Article Snippet: Four commercially available kits for these four proteins; angiogenin (ANG) [Abcam, UK - ab99970], clusterin (CLU) [R&D system, UK - DCLU00], C-C Motif Chemokine 18 (CCL18) [Abcam, UK - ab100620], and Complement C1q [Abcam, UK - ab170246] were used.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: BBA Clinical

Article Title: Novel panel of protein biomarkers to predict response to bortezomib-containing induction regimens in multiple myeloma patients

doi: 10.1016/j.bbacli.2017.05.003

Figure Lengend Snippet: Logistic regression analysis data. List of different protein combinations used to establish the best model that can be used as a predictive panel for response to induction therapy containing bortezomib regime.

Article Snippet: Four commercially available kits for these four proteins; angiogenin (ANG) [Abcam, UK - ab99970], clusterin (CLU) [R&D system, UK - DCLU00], C-C Motif Chemokine 18 (CCL18) [Abcam, UK - ab100620], and Complement C1q [Abcam, UK - ab170246] were used.

Techniques:

Journal: Molecular Metabolism

Article Title: LRP1 in GABAergic neurons is a key link between obesity and memory function

doi: 10.1016/j.molmet.2024.101941

Figure Lengend Snippet: Selective deletion of LRP1 in GABAergic neurons causes metabolic disorders. ( A ) Body weight, ( B ) epididymal fat, ( C ) serum leptin, ( D ) serum FFA, ( E ) serum ApoJ, ( F ) blood glucose, ( G ) serum insulin, and ( H ) HOMA-IR were measured in LRP1 loxP/loxP and Vgat-Cre; LRP1 loxP/loxP male mice that were overnight fasted mice at 32 weeks of age. n = 12 for control, n = 18 for Vgat-Cre; LRP1 loxP/loxP . ( I ) Body weight, ( J ) blood glucose, ( K ) serum insulin, ( L ) serum leptin, and ( M ) serum ApoJ were measured in LRP1 loxP/loxP and Vgat-Cre; LRP1 loxP/loxP male mice that were overnight fasted at 16 weeks of age. n = 12 for control, n = 7 for Vgat-Cre; LRP1 loxP/loxP . All graphs represent means or individual values ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. LRP1 loxP/loxP by two-sided Student's t-test.

Article Snippet: Serum levels of insulin (90080, Crystal Chem, Chicago, IL), leptin (90030, Crystal Chem, Chicago, IL), apolipoprotein J (ApoJ) (MCLU00, R&D Systems, Minneapolis, MN), Aβ 40 (KMB3481, Thermo Fisher Scientific Inc., Waltham, MA), Aβ 42 (KMB3441, Thermo Fisher Scientific Inc., Waltham, MA) and free fatty acids (FFA) (ab65341, Abcam, Waltham, MA) were measured by enzyme-linked immunosorbent assay (ELISA).

Techniques: Control

Journal: Molecular Metabolism

Article Title: LRP1 in GABAergic neurons is a key link between obesity and memory function

doi: 10.1016/j.molmet.2024.101941

Figure Lengend Snippet: Relationship of motor function, motor coordination, and spatial recognition memory with obesity-linked metabolic parameters. Correlation of body weight with ( A ) total distance traveled on the locomotion test, ( B ) latency to fall on the rotarod test, ( C ) total distance traveled, and ( D ) time spent in the novel arm on the spatial Y-maze test was evaluated. Correlation of serum insulin with ( E ) latency to fall on the rotarod test, ( F ) total distance traveled, and ( G ) time spent in the novel arm on the spatial Y-maze test was evaluated. Correlation of serum leptin with ( H ) latency to fall on the rotarod test, ( I ) total distance traveled, and ( J ) time spent in the novel arm on the spatial Y-maze test was evaluated. Correlation of serum ApoJ with ( K ) latency to fall on the rotarod test and (L) time spent in the novel arm on the spatial Y-maze test were evaluated. The P values were obtained by Pearson correlation analysis, and the r values indicate Pearson correlation coefficient. All correlations were analyzed from the data of LRP1 loxP/loxP and Vgat-Cre; LRP1 loxP/loxP male mice (32 weeks old).

Article Snippet: Serum levels of insulin (90080, Crystal Chem, Chicago, IL), leptin (90030, Crystal Chem, Chicago, IL), apolipoprotein J (ApoJ) (MCLU00, R&D Systems, Minneapolis, MN), Aβ 40 (KMB3481, Thermo Fisher Scientific Inc., Waltham, MA), Aβ 42 (KMB3441, Thermo Fisher Scientific Inc., Waltham, MA) and free fatty acids (FFA) (ab65341, Abcam, Waltham, MA) were measured by enzyme-linked immunosorbent assay (ELISA).

Techniques:

Journal: Molecular Metabolism

Article Title: LRP1 in GABAergic neurons is a key link between obesity and memory function

doi: 10.1016/j.molmet.2024.101941

Figure Lengend Snippet: Relationship between metabolic parameters and cognitive function. Correlation of body weight, serum leptin, serum insulin, and serum ApoJ with the data from acquisition ( A, C, E, G ) and reversal ( B, D, F, H ) water T-maze on day 1 was evaluated. The P values were obtained by Pearson correlation analysis and r values indicate the Pearson correlation coefficient. All correlations were analyzed from the data of LRP1 loxP/loxP and Vgat-Cre; LRP1 loxP/loxP male mice (32 weeks old).

Article Snippet: Serum levels of insulin (90080, Crystal Chem, Chicago, IL), leptin (90030, Crystal Chem, Chicago, IL), apolipoprotein J (ApoJ) (MCLU00, R&D Systems, Minneapolis, MN), Aβ 40 (KMB3481, Thermo Fisher Scientific Inc., Waltham, MA), Aβ 42 (KMB3441, Thermo Fisher Scientific Inc., Waltham, MA) and free fatty acids (FFA) (ab65341, Abcam, Waltham, MA) were measured by enzyme-linked immunosorbent assay (ELISA).

Techniques:

Journal: OncoImmunology

Article Title: Aberrant fucosylation enables breast cancer clusterin to interact with dendritic cell-specific ICAM-grabbing non-integrin (DC-SIGN)

doi: 10.1080/2162402x.2019.1629257

Figure Lengend Snippet: Figure 1. Clusterin expression in luminal breast cancer samples. A and B. Clusterin expression was analyzed by immunohistochemistry in tumoral (a) and juxtatumoral (b) tissues. Representative pictures are shown (n = 3, bars: 50 µm). C. Clusterin expression was quantified in tumor samples and their juxtatumoral counterparts by ELISA. Results are expressed as the amount of clusterin/total protein (µg of clusterin/mg of total protein, n = 21, ns = no statistically significant differences, p = .31).

Article Snippet: Total clusterin was detected by ELISA using a conventional ELISA kit (Human Clusterin DuoSet ELISA, R&D Systems).

Techniques: Expressing, Immunohistochemistry, Enzyme-linked Immunosorbent Assay

Journal: OncoImmunology

Article Title: Aberrant fucosylation enables breast cancer clusterin to interact with dendritic cell-specific ICAM-grabbing non-integrin (DC-SIGN)

doi: 10.1080/2162402x.2019.1629257

Figure Lengend Snippet: Figure 2. Fucosylated clusterin (fCLU) is expressed in luminal breast cancer and interacts with DC-SIGN. a. Scheme of the ELISA assay used to detect fucosylated clusterin. Clusterin is captured by a mAb directed to clusterin and the presence of fucosylated clusterin (fCLU) is revealed by using either biotinilated Ulex europaeus agglutinin-I lectin (UEA-I) or Lotus tetragonolobus lectin (LT). b. Fucosylated semen clusterin, but not serum or recombinant clusterin (blue bars), was detected by the assay described in (a). The addition of α-L-fucose prevents the recognition of fucosylated semen clusterin (red bars). Results are expressed as the mean ± SD of 4 independent experiments. c and d. Fucosylated clusterin expression in breast tumor and juxtatumoral samples (n = 14–21) was analyzed using Lotus tetragonolobus lectin (c) or Ulex europaeus agglutinin-I (d). Results are expressed as the amount of fucosylated clusterin relative to the amount of total clusterin (left panels) or total protein (right panels) (***p < .001, **p < .01, *p < .05). E. The ability of clusterin from tumor samples and juxtatumoral samples to bind to DC-SIGN was evaluated. Upper panel: the presence of clusterin was evaluated using a mAb directed to clusterin. Lower panel: the membrane was stripped and revealed using DC-SIGN-huFc and HRP conjugated anti-huIgG. Of note, total clusterin but not total protein was used to normalize the amount of sample loaded into the gel for each tumor and juxtatumor sample pair. Accordingly, no loading control was analyzed. Panel F shows the ratio between DC-SIGN-huFC binding and total CLU. The fold change between tumor and juxtatumor CLU ratios are shown.

Article Snippet: Total clusterin was detected by ELISA using a conventional ELISA kit (Human Clusterin DuoSet ELISA, R&D Systems).

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Expressing, Membrane, Control, Binding Assay

Journal: OncoImmunology

Article Title: Aberrant fucosylation enables breast cancer clusterin to interact with dendritic cell-specific ICAM-grabbing non-integrin (DC-SIGN)

doi: 10.1080/2162402x.2019.1629257

Figure Lengend Snippet: Figure 6. Fucosylated clusterin promotes the differentiation of macrophages into a proangiogenic profile. a-c. Macrophages were differentiated from monocytes cultured with M-CSF for 5 days, in the absence or presence of semen fucosylated clusterin. Then, cells were stimulated, or not, with LPS (10 ng/ml), and the pattern of cell clustering (a), the expression of HLA-DR, CD86, PDL-1, CD40 and CD80 (b), and the production of VEGF, IL-8, TNF-α, IL-10, and IL-6 (c) were evaluated by optical microscopy, flow cytometry or ELISA. Representative pictures (bars = 100µm) and histograms (n = 3–9) are shown in a and b. On b, the control condition mean fluorescent intensities (MFI) of each independent experiment were normalized as 1. The mean of 5–9 independent experiments is shown. The mean of 5–7 independent experiments is shown in c (**p < .01, *p < .05).

Article Snippet: Total clusterin was detected by ELISA using a conventional ELISA kit (Human Clusterin DuoSet ELISA, R&D Systems).

Techniques: Cell Culture, Expressing, Microscopy, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Control

Journal: OncoImmunology

Article Title: Aberrant fucosylation enables breast cancer clusterin to interact with dendritic cell-specific ICAM-grabbing non-integrin (DC-SIGN)

doi: 10.1080/2162402x.2019.1629257

Figure Lengend Snippet: Figure 5. Fucosylated clusterin interacts with macrophages. a. Expression of DC-SIGN by monocyte-derived macrophages. A representative experiment (n = 7) is shown. b. Macrophages were incubated for 30 min at 4°C with different concentrations of fucosylated clusterin isolated from semen. Cells were then washed, lysed and the binding of clusterin was revealed by ELISA. Results are expressed as the mean ± SD of 4 independent experiments. c. Macrophages were incubated for 30 min at 4°C with fucosylated clusterin (10 µg/ml), in the absence or presence of a neutralizing mAb directed to DC- SIGN (5 µg/ml), lactose (100 µg/ml), sucrose (100 µg/ml), mannan (1–100 µg/ml), or α-L-fucose (1–100 µg/ml). Cells were then washed, lysed and the binding of clusterin was revealed by ELISA. Results are expressed as the mean ± SD of 3 experiments (**p < .01, *p < .05). d. Macrophages were incubated for 15 min at 37°C with fucosylated clusterin (10 µg/ml) and clusterin endocytosis (green) was analyzed by confocal microscopy (n = 5, bars:10µm).

Article Snippet: Total clusterin was detected by ELISA using a conventional ELISA kit (Human Clusterin DuoSet ELISA, R&D Systems).

Techniques: Expressing, Derivative Assay, Incubation, Isolation, Binding Assay, Enzyme-linked Immunosorbent Assay, Confocal Microscopy

Journal: The Journal of Clinical Investigation

Article Title: Schlafen 4–expressing myeloid-derived suppressor cells are induced during murine gastric metaplasia

doi: 10.1172/JCI82529

Figure Lengend Snippet: (A) Serum SHH levels in WT and pCMV-Shh mice infected with H. felis over 6 months. n = 5–10 mice per time point over 3 experiments. One-way ANOVA followed by Dunnett’s multiple comparisons test on log-transformed values was performed. P values are relative to UI; ‡P < 0.001, #P < 0.0001. (B) Representative H&E images of gastric mucosa from WT and pCMV-Shh mice at the indicated times. Scale bars: 100 μm. Histologic scoring of gastric (C) PMN infiltration and (D) metaplasia for n = 8–10 mice per time point over 3 experiments. Kruskal-Wallis ANOVA with Dunn’s test of multiple comparisons was performed. *P < 0.05; horizontal lines represent the median and interquartile range. (E) Images of 4-month-infected gastric corpus stained with GSII lectin (green), clusterin (CLU, green), GIF (red), and H+-K+-ATPase (HK, white) in WT and pCMV-Shh mice. Scale bars: 50 μm. n = 5 mice per group for B and E.

Article Snippet: For immunofluorescence staining, slides were incubated with the following antibodies overnight: GSII-FITC (1:1,000, FL-1211, Vector Laboratories), arginase I (1:100, sc-20150, Santa Cruz Biotechnology Inc.), IFN-β (1:100, AP30401PU-N, Acris Antibodies), IFN-α–FITC (1:200, 22100-3, Interferon Source), H + -K + -ATPase-β (1:1,000, D032-3, Medical and Biological Laboratories), caspase-3 (1:400, 9661, Cell Signaling Technology), clusterin (1:2,000, sc-6420, Santa Cruz Biotechnology Inc.), E-cadherin (1:1,000, sc-7870, Santa Cruz Biotechnology Inc.), CD15 (1:7,500, 13-0159-1631, eBioscience), SLFN12L (1:100, NBP1-91060, Novus).

Techniques: Infection, Transformation Assay, Staining

Journal: Acta Neuropathologica Communications

Article Title: Clusterin ameliorates tau pathology in vivo by inhibiting fibril formation

doi: 10.1186/s40478-020-01079-1

Figure Lengend Snippet: Clusterin co-localizes with tau deposits and is upregulated in human tauopathies. a Human brain tissues representing normal control, Alzheimer’s disease (AD), and the primary tauopathies Pick’s disease (PiD) and corticobasal degeneration (CBD). Co-localization of CLU (brown) with tau deposits, marked by MC-1 labeling (blue). Scale bar, 100 μm. b Arrows indicate co-localization of CLU (red) with mature tau tangles labeled by thioflavine-S staining (green). Arrowheads show tau tangles without CLU co-localization. Asterisks represent amyloid plaques. Scale bar, 100 μm. c Biochemical evaluation of the total CLU protein levels in the cortical region of human tauopathies. N = 10–15 cases/group. Data presented as mean ± S.E.M. and analyzed with one-way ANOVA with Tukey’s multiple comparison test, * p < 0.05, ** p < 0.01

Article Snippet: Mouse CLU levels were measured using the Mouse Clusterin DuoSet ELISA (R&D Systems, DY2747), according to manufacturer’s instructions.

Techniques: Control, Labeling, Staining, Comparison

Journal: Acta Neuropathologica Communications

Article Title: Clusterin ameliorates tau pathology in vivo by inhibiting fibril formation

doi: 10.1186/s40478-020-01079-1

Figure Lengend Snippet: Loss of Clusterin leads to behavioral abnormalities. a - b Behavioral and cognitive abilities were assessed using a the open field test (OFA) and b elevated plus maze (EPM) in CLU WT-GFP (N = 49), CLU KO-GFP (N = 12), CLU WT-Tau P301L (N = 65), and CLU KO-Tau P301L (N = 23). OFA was used to test anxiety-related behavior, evaluated by time spent in the center compared to total time travelled. EPM measured the time spent in the open arms during the test which is a reflection of exploratory behavior. Data present as mean ± S.E.M. and analyzed with two-way ANOVA with Fisher’s LSD test, * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Mouse CLU levels were measured using the Mouse Clusterin DuoSet ELISA (R&D Systems, DY2747), according to manufacturer’s instructions.

Techniques:

Journal: Acta Neuropathologica Communications

Article Title: Clusterin ameliorates tau pathology in vivo by inhibiting fibril formation

doi: 10.1186/s40478-020-01079-1

Figure Lengend Snippet: Clusterin reduces the severity of tau pathology. a Representative images from the cortical and hippocampal regions of 6-month-old CLU WT-Tau P301L and CLU KO-Tau P301L showing tau pathology. Tau hyperphosphorylation at serine 202, serines 396/404, and tau conformational change were detected by using CP-13, PHF-1, and MC-1, respectively. Scale bar, 100 μm. b Quantitative analysis of tau accumulation in cortex and hippocampus of CLU WT-Tau P301L and CLU KO-Tau P301L mice. For the CP-13, PHF-1 and MC-1 analyses n = 23–28 mice/group were used. Data presented as mean ± S.E.M. and analyzed with Student’s t test, * p < 0.05. c Biochemical evaluation of the total tau levels in the TBS-soluble (S1) and sarkosyl-soluble (S2) fractions. N = 22–25 mice/group. Data present as mean ± S.E.M. and analyzed with Student’s t test, * p < 0.05. d The levels of total and hyperphosphorylated tau were assessed by the immunoblotting analysis. N = 12 mice/group. Data present as mean ± S.E.M. and analyzed with Student’s t test, * p < 0.05

Article Snippet: Mouse CLU levels were measured using the Mouse Clusterin DuoSet ELISA (R&D Systems, DY2747), according to manufacturer’s instructions.

Techniques: Western Blot