Journal: BMB Reports

Article Title: SPINK1 promotes cell growth and metastasis of lung adenocarcinoma and acts as a novel prognostic biomarker

doi: 10.5483/BMBRep.2018.51.12.205

Figure Lengend Snippet: SPINK1 enhances the migration and invasion of LAC cells by up-regulating MMP12. (A) MMPs expressions were analyzed by qRT-PCR in A549 cells and H1975 cells transfected with scramble or SPINK1 siRNAs. (B) MMP12 protein expressions were analyzed by Western blot in A549 cells and H1975 cells transfected with scramble or SPINK1 siRNA 1. (C) MMP12 protein levels were measured by Western blot in PC9 cells and H1299 cells transfected with control or SPINK1. (D) MMP12 mRNA expression in xenograft tissues was measured by qPCR. (E) The correlation between SPINK1 expression and MMP12 expression in LAC tissues was analyzed with Pearson correlation. (F) The interference effect of MMP12 siRNA was verified by Western blot in PC9 and H1299 cells stably overexpressing SPINK1. The number of migrating and invasive cells was counted (G). (H) Recombinant human MMP12 (rhMMP12) or control IgG (cIgG) at a concentration of 50 ng/ml was added back into SPINK1 knockdown A549 and H1299 cells, and the number of migrating and invasive cells was counted. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Recombinant human MMP12 (rhMMP12, 917-MPB) and the control IgG (cIgG, 1-001-A) were purchased from R&D Systems.

Techniques: Migration, Quantitative RT-PCR, Transfection, Western Blot, Control, Expressing, Stable Transfection, Recombinant, Concentration Assay, Knockdown

Journal: Cardiovascular Diabetology

Article Title: Genetic deletion of MMP12 ameliorates cardiometabolic disease by improving insulin sensitivity, systemic inflammation, and atherosclerotic features in mice

doi: 10.1186/s12933-023-02064-3

Figure Lengend Snippet: Correlations of circulating MMP12 with clinical and laboratory indicators for cardiometabolic diseases (A) Violin plots of MMP12 concentrations in serum from healthy volunteers (HV) and from patients with metabolic syndrome (MS). Groups were compared using the Mann-Whitney U test (n = 65 − 63). (B) Receiver operating characteristic (ROC) curve for the prediction of MS using MMP12. The graph shows the ROC curve, area under the curve (AUC), 95% confidence interval, and the best cut-off value according to Youden index along with specificity and sensitivity. (C) Heat map for correlation analyses of serum MMP12 concentrations with body mass index (BMI), as well as serum levels of glucose, lipoprotein (a) (Lp(a)), interleukin 6 (IL-6), and bilirubin levels. Correlations were quantified using Spearman’s correlation coefficient (r). Correlation plots of circulating MMP12 concentrations (pg/ml) with (D) BMI (kg/m 2 ), (E) glucose (mg/dl), (F) Lp(a) (mg/dl), (G) IL-6 (mg/dl), and (H) bilirubin (mg/dl) (n = 128)

Article Snippet: Serum MMP12 concentrations were measured using a quantitative sandwich enzyme immunoassay (#EK0950; Boster Bio, Pleasanton, CA) according to the manufacturer’s instruction and negative values were set to zero.

Techniques: MANN-WHITNEY

Journal: BMC Pulmonary Medicine

Article Title: Optimization of total protein and activity assays for the detection of MMP-12 in induced human sputum

doi: 10.1186/1471-2466-10-40

Figure Lengend Snippet: A comparison of commercially available MMP-12 antibodies as capture antibodies in the FRET activity assay format .

Article Snippet: MMP-12 protein from R&D Systems was autocatalyzed and spiked into four human sputum samples and into assay diluent at three different concentrations (final theoretical spiking concentrations of 10,000, 3000, and 1000 pg/mL).

Techniques: Comparison, Activity Assay

Journal: BMC Pulmonary Medicine

Article Title: Optimization of total protein and activity assays for the detection of MMP-12 in induced human sputum

doi: 10.1186/1471-2466-10-40

Figure Lengend Snippet: A plot of RFU (abscissa) vs. analyte (ordinate) in the capture FRET format using antibodies Dendritics 282 (cat# DDX0282), Abcam 11614, or Genway 066 (cat# 15-288-066A) . Analytes are present at 1 ug/mL, except for 12a - autocatalyzed R&D Systems MMP-12 at 1 ng/mL, 12 Wyeth cat - recombinant catalytic MMP-12 at 10 ng/mL, 12 pro - pro form MMP-12, R&D Systems and 12 autocat - autocatalyzed R&D Systems MMP-12 both at 10 ng/mL. n171 and n168 are two sputum samples with unknown MMP-12 concentrations.

Article Snippet: MMP-12 protein from R&D Systems was autocatalyzed and spiked into four human sputum samples and into assay diluent at three different concentrations (final theoretical spiking concentrations of 10,000, 3000, and 1000 pg/mL).

Techniques: Recombinant

Journal: BMC Pulmonary Medicine

Article Title: Optimization of total protein and activity assays for the detection of MMP-12 in induced human sputum

doi: 10.1186/1471-2466-10-40

Figure Lengend Snippet: Vmax (abscissa) is plotted against Anaspec substrate (ordinate) in a solution phase assay using 20 μM substrate and 100 ng/mL pro or autocatalyzed MMP-12 .

Article Snippet: MMP-12 protein from R&D Systems was autocatalyzed and spiked into four human sputum samples and into assay diluent at three different concentrations (final theoretical spiking concentrations of 10,000, 3000, and 1000 pg/mL).

Techniques:

Journal: BMC Pulmonary Medicine

Article Title: Optimization of total protein and activity assays for the detection of MMP-12 in induced human sputum

doi: 10.1186/1471-2466-10-40

Figure Lengend Snippet: A pairwise plot of interpolated MMP-12 concentrations for both activity and total MMP-12 protein concentration in normal, COPD, and asthmatic donors .

Article Snippet: MMP-12 protein from R&D Systems was autocatalyzed and spiked into four human sputum samples and into assay diluent at three different concentrations (final theoretical spiking concentrations of 10,000, 3000, and 1000 pg/mL).

Techniques: Activity Assay, Protein Concentration

Journal: BMC Pulmonary Medicine

Article Title: Optimization of total protein and activity assays for the detection of MMP-12 in induced human sputum

doi: 10.1186/1471-2466-10-40

Figure Lengend Snippet: A boxplot of condition (abscissa) vs interpolated MMP-12 protein concentration (ordinate) trellised by assay format .

Article Snippet: MMP-12 protein from R&D Systems was autocatalyzed and spiked into four human sputum samples and into assay diluent at three different concentrations (final theoretical spiking concentrations of 10,000, 3000, and 1000 pg/mL).

Techniques: Protein Concentration

Journal: PLoS Medicine

Article Title: An Immune Basis for Lung Parenchymal Destruction in Chronic Obstructive Pulmonary Disease and Emphysema

doi: 10.1371/journal.pmed.0010008

Figure Lengend Snippet: (A) CD14 + , lymphocyte-depleted lung leukocytes were cultured with and without the indicated amounts of recombinant human IP-10 and IFN-γ, and supernatants were assessed for the presence of MMP12 by Western blotting. (B) Fold increase relative to unstimulated of MMP12 mRNA from lung macrophages stimulated without ( – ) and with ( + ) 500 ng/ml of IP-10 in the presence or absence of a function-blocking antibody to CXCR3 as determined by real-time PCR. (C and D) Lung tissue from a participant with emphysema (C) shows strong immune staining for MMP12 localized to macrophages (arrows), and (D) shows lung tissue from a control participant without emphysema and with undetectable MMP12. The insets show a high-power view of lung macrophages staining positive (C) and negative (D) for MMP12 (×60) *, p = 0.04.

Article Snippet: Supernatants were collected, and MMP12 was detected using anti-human MMP12 (R&D Systems) by Western blotting according to the manufacturer's instructions.

Techniques: Cell Culture, Recombinant, Western Blot, Blocking Assay, Real-time Polymerase Chain Reaction, Staining, Control