Journal: International Journal of Molecular Sciences

Article Title: Elevation of Tear MMP-9 Concentration as a Biomarker of Inflammation in Ocular Pathology by Antibody Microarray Immunodetection Assays

doi: 10.3390/ijms23105639

Figure Lengend Snippet: Detection and quantification of MMP-9 enzyme inflammation biomarker in human tear samples using AbMAs. First, the selected antibodies were immobilized onto glass slides that were incubated with the sample. Then, MMP-9 was captured by the mentioned antibody and detected with a labeled antibody cocktail. Finally, the intensity of the signal was quantified and the data acquired, allowing the analysis of the MMP-9 biomarker in the samples.

Article Snippet: Each AbMA had two replicate spots of rabbit IgG anti-human MMP-9 (#10327-R043, Sino Biological, Beijing, China) immobilized at 200 µg/mL onto SIVG printing solution at 0.05% (IMG Pharma Biotech S.L., Derio, Spain).

Techniques: Biomarker Assay, Incubation, Labeling

Journal: International Journal of Molecular Sciences

Article Title: Elevation of Tear MMP-9 Concentration as a Biomarker of Inflammation in Ocular Pathology by Antibody Microarray Immunodetection Assays

doi: 10.3390/ijms23105639

Figure Lengend Snippet: Concentration of MMP-9 in the collection of tear samples from healthy controls, without any ocular disorder diagnosed, using ELISA (blue) as the gold standard technique and AbMAs (orange). MMP-9 concentration is represented as ng/mL for each individual. A gray line is plotted at 30 ng/mL of MMP-9 enzyme in tear, the threshold value at which higher concentrations are considered a sign of ocular inflammation.

Article Snippet: Each AbMA had two replicate spots of rabbit IgG anti-human MMP-9 (#10327-R043, Sino Biological, Beijing, China) immobilized at 200 µg/mL onto SIVG printing solution at 0.05% (IMG Pharma Biotech S.L., Derio, Spain).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Elevation of Tear MMP-9 Concentration as a Biomarker of Inflammation in Ocular Pathology by Antibody Microarray Immunodetection Assays

doi: 10.3390/ijms23105639

Figure Lengend Snippet: ng/mL of MMP-9 in ocular inflammation patient tear samples quantified using AbMA. A gray line is plotted at 30 ng/mL of MMP-9 enzyme in tear, the threshold value at which higher concentrations are considered a sign of ocular inflammation.

Article Snippet: Each AbMA had two replicate spots of rabbit IgG anti-human MMP-9 (#10327-R043, Sino Biological, Beijing, China) immobilized at 200 µg/mL onto SIVG printing solution at 0.05% (IMG Pharma Biotech S.L., Derio, Spain).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Elevation of Tear MMP-9 Concentration as a Biomarker of Inflammation in Ocular Pathology by Antibody Microarray Immunodetection Assays

doi: 10.3390/ijms23105639

Figure Lengend Snippet: Tear MMP-9 concentration differences in the groups of patients suffering ocular inflammation versus the group of healthy controls. Cliff’s delta values are displayed for each comparison. ( A ) Differences between healthy controls and MGD, DE, and allergy patients (other pathologies group). ( B ) Differences between healthy controls and cataracts patients. ( C ) Differences between healthy controls and glaucoma patients.

Article Snippet: Each AbMA had two replicate spots of rabbit IgG anti-human MMP-9 (#10327-R043, Sino Biological, Beijing, China) immobilized at 200 µg/mL onto SIVG printing solution at 0.05% (IMG Pharma Biotech S.L., Derio, Spain).

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Elevation of Tear MMP-9 Concentration as a Biomarker of Inflammation in Ocular Pathology by Antibody Microarray Immunodetection Assays

doi: 10.3390/ijms23105639

Figure Lengend Snippet: Confusion matrix of tear MMP-9 analysis over the different samples. True positive (TP), false positive (FP), false negative (FN), and true negative (TN) rates are detailed. Sensitivity is calculated as TP/(TP + FN), specificity as TN/(TN + FP), and accuracy as (TP + TN)/(TP + FP + FN + TN).

Article Snippet: Each AbMA had two replicate spots of rabbit IgG anti-human MMP-9 (#10327-R043, Sino Biological, Beijing, China) immobilized at 200 µg/mL onto SIVG printing solution at 0.05% (IMG Pharma Biotech S.L., Derio, Spain).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Elevation of Tear MMP-9 Concentration as a Biomarker of Inflammation in Ocular Pathology by Antibody Microarray Immunodetection Assays

doi: 10.3390/ijms23105639

Figure Lengend Snippet: Schematic representation of a microscope glass slide with AbMAs printed. Twenty-four AbMAs with two spots of rabbit IgG anti-human MMP-9 at 200 µg/mL in SIVG 0.05% were immobilized onto treated slides. Image created with BioRender.com.

Article Snippet: Each AbMA had two replicate spots of rabbit IgG anti-human MMP-9 (#10327-R043, Sino Biological, Beijing, China) immobilized at 200 µg/mL onto SIVG printing solution at 0.05% (IMG Pharma Biotech S.L., Derio, Spain).

Techniques: Microscopy

Journal: Journal of Cellular and Molecular Medicine

Article Title: Relationship among LRP1 expression, Pyk2 phosphorylation and MMP‐9 activation in left ventricular remodelling after myocardial infarction

doi: 10.1111/jcmm.13113

Figure Lengend Snippet: Primary antibodies used for immunohistochemical analysis

Article Snippet: MMP‐9 , TP221 , AMS Biotechnology.

Techniques: Immunohistochemical staining

Journal: bioRxiv

Article Title: Ribosomal protein L23 drives the metastasis of hepatocellular carcinoma via upregulating MMP9

doi: 10.1101/2021.07.27.453993

Figure Lengend Snippet: (A, B) EMT-related markers (MMP9, MMP2, N-cadherin, Vimentin, Smad2, Twist1 and E-cadherin) were measured on RPL23-depleted HCC cells by qRT-PCR. β-actin was used as an internal quantitative control. (***p<0.001) (C) RPL23 regulated MMP9 protein expression in HLE and MHCC97H cells measured by western blot assay. GAPDH was used as a loading control for western blotting. (D) The half-life of MMP-9 mRNA was reduced after RPL23 knockdown in HLE and MHCC97H cells followed by treatment with 5ug/mL actinomycin D at the indicated times. Error bars represent SEM. p-values (HLE): **p = 0.00116 (shCont vs shRPL23#1), **p = 0.00296 (shCont vs shRPL23#2). p-values (MHCC97H): **p = 0.00314 (shCont vs shRPL23#1), **p = 0.00477 (shCont vs shRPL23#2). (E) RIP assays showed that RPL23 directly bound to MMP9 mRNA. (F) RNA pull-down results showed that RPL23 was directly associated with the 3’UTR of MMP9 mRNA. Control indicates a control pulldown containing beads only. Representative data are from at least three independent experiments. Data are shown as mean ± SD. *p<0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The primary antibodies were as follows: rabbit anti-RPL23 (Proteintech, 16086-1-AP), rabbit anti-MMP9 (OriGene, TA326652), anti-GAPDH (Santa Cruz Biotechnology, sc-365062).

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: bioRxiv

Article Title: Ribosomal protein L23 drives the metastasis of hepatocellular carcinoma via upregulating MMP9

doi: 10.1101/2021.07.27.453993

Figure Lengend Snippet: (A) The protein level of MMP9 increased after overexpression of RPL23 in Huh7 cells. The half-life of MMP-9 mRNA was increased after RPL23 expression in Huh7 cells followed by treatment with 5ug/mL actinomycin D at the indicated times. *p = 0.0187 (Vector vs RPL23). Representative data are from at least three independent experiments. Data are shown as mean ± SD.*p<0.05.

Article Snippet: The primary antibodies were as follows: rabbit anti-RPL23 (Proteintech, 16086-1-AP), rabbit anti-MMP9 (OriGene, TA326652), anti-GAPDH (Santa Cruz Biotechnology, sc-365062).

Techniques: Over Expression, Expressing, Plasmid Preparation

Journal: bioRxiv

Article Title: Ribosomal protein L23 drives the metastasis of hepatocellular carcinoma via upregulating MMP9

doi: 10.1101/2021.07.27.453993

Figure Lengend Snippet: (A, B) Overexpression of MMP9 rescued the inhibition effect of decreased RPL23 on HCC cell proliferation. (C) overexpression of MMP-9 rescued the repression effect of knockdown RPL23 on HCC cell migration ability by wound-healing assay. The cells were counted from 6 images. (D) Upregulation of MMP-9 could significantly rescued the effects of decreased RPL23 in HLE and MHCC97H cells for both migration and invasion by transwell assays. The cells were counted from 5 images. Representative data are from at least three independent experiments. Data are shown as mean ± SD. **P < 0.01, ***P < 0.001.

Article Snippet: The primary antibodies were as follows: rabbit anti-RPL23 (Proteintech, 16086-1-AP), rabbit anti-MMP9 (OriGene, TA326652), anti-GAPDH (Santa Cruz Biotechnology, sc-365062).

Techniques: Over Expression, Inhibition, Migration, Wound Healing Assay

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer

doi: 10.3390/ijms241914847

Figure Lengend Snippet: MMP9 and miR-21 gene editing with CRISPR-Cas9. ( A ) DNA sequencing of the PX-330 plasmid with the sequence inserts for MMP9 (sgRNA1 and sgRNA2) at the beginning of MMP9 Exon 1 on chromosome 20q ( left ) and DNA sequencing of the sequence inserts for miR-21 (sgRNA 1 and sgRNA2), located in two regions of chromosome 17 ( right ). ( B ) miR-21 gene expression in samples edited with miR-21 sgRNA 1 and their respective control transfected with the plasmid without any insert (Scramble) in the PC-3 cell line. ( C , D ) MMP9 and RECK gene expression in samples edited with CRISPR-Cas9 MMP9, sgRNAs 1 and 2 or miR-21 sgRNA1 compared with the scramble control in the PC-3 cell line. ( E ) miR-21 gene expression in samples edited with CRISPR-Cas9 miR-21 sgRNA 1 compared to the scramble control in the DU145 cell line. ( F , G ) MMP9 and RECK gene expression in samples edited with CRISPR-Cas9 MMP9 sgRNAs 1 and 2 or miR-21 sgRNA1 compared to Scramble in the DU145 cell line. Statistical significance set at p < 0.05.

Article Snippet: Cells were fixed in 4% paraformaldehyde in PBS for 10 min at room temperature, washed 3 times with PBS and subjected to blocking solution with 3% BSA in PBS for 1 h. The MMP9 (1:150, Boster M00139) and RECK (1:100, Santa Cruz SC373929) antibodies were incubated in PBS containing 1% BSA overnight at 4 °C.

Techniques: CRISPR, DNA Sequencing, Plasmid Preparation, Sequencing, Gene Expression, Control, Transfection

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer

doi: 10.3390/ijms241914847

Figure Lengend Snippet: MMP9 and RECK protein expression in MMP9 and miR-21 CRISPR-Cas9-edited metastatic PCa cell lines. ( A ) Western blot analysis of MMP9 protein content in PC-3 and DU145 cells edited with MMP9 sgRNA 1/2 or miR-21 sgRNA1. ( B , C ) Protein immunofluorescence of colocalized MMP9 (green) and RECK (red) in samples edited with MMP9 sgRNA 1/2 or miR-21 sgRNA1 in both PC-3 and DU145 cells.

Article Snippet: Cells were fixed in 4% paraformaldehyde in PBS for 10 min at room temperature, washed 3 times with PBS and subjected to blocking solution with 3% BSA in PBS for 1 h. The MMP9 (1:150, Boster M00139) and RECK (1:100, Santa Cruz SC373929) antibodies were incubated in PBS containing 1% BSA overnight at 4 °C.

Techniques: Expressing, CRISPR, Western Blot, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer

doi: 10.3390/ijms241914847

Figure Lengend Snippet: Gene expression of miR-21 targets in metastatic PCa cell lines. Gene expression of ( A ) MARKS, ( B ) BTG2, and ( C ) PDCD4 in MMP9 and miR-21 CRISPR-Cas9-edited PC-3 cells, and ( D ) MARKS, ( E ) BTG2, and ( F ) PDCD4 in MMP9 and miR-21 CRISPR-Cas9-edited DU145 cells.

Article Snippet: Cells were fixed in 4% paraformaldehyde in PBS for 10 min at room temperature, washed 3 times with PBS and subjected to blocking solution with 3% BSA in PBS for 1 h. The MMP9 (1:150, Boster M00139) and RECK (1:100, Santa Cruz SC373929) antibodies were incubated in PBS containing 1% BSA overnight at 4 °C.

Techniques: Gene Expression, CRISPR

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer

doi: 10.3390/ijms241914847

Figure Lengend Snippet: Gene expression of CDH1, integrins, BAX, and mTOR in metastatic PCa lines. Gene expression of ( A ) CDH1 cadherin, integrins ( B ) ITGB3 and ( C ) ITGB1, ( D ) BAX and ( E ) mTOR in MMP9 sgRNA 1/2- and miR-21 sgRNA1 CRISPR-Cas9-edited PC-3 cells. Gene expression of ( F ) CDH1 cadherin, integrins ( G ) ITGB3 and ( H ) ITGB1, ( I ) BAX and ( J ) mTOR in MMP9 sgRNA 1/2- and miR-21 sgRNA1 CRISPR-Cas9-edited DU145 cell line.

Article Snippet: Cells were fixed in 4% paraformaldehyde in PBS for 10 min at room temperature, washed 3 times with PBS and subjected to blocking solution with 3% BSA in PBS for 1 h. The MMP9 (1:150, Boster M00139) and RECK (1:100, Santa Cruz SC373929) antibodies were incubated in PBS containing 1% BSA overnight at 4 °C.

Techniques: Gene Expression, CRISPR

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer

doi: 10.3390/ijms241914847

Figure Lengend Snippet: Flow cytometry for assessing proliferation and apoptosis in the metastatic PC-3 and DU145 cell lines. ( A ) Labeling MMP9 sgRNA1/2- and miR-21 sgRNA1 CRISPR-Cas9-edited PC-3 cells with a ki67 antibody to evaluate cell proliferation rate. ( B – D ) Labeling MMP9 sgRNA1/2 and miR-21 sgRNA1 CRISPR-Cas9-edited PC-3 cells with annexin-5 and 7-AAD to calculate the percentage of cells in the early and late stages of apoptosis and total apoptosis. ( E ) Labeling MMP9 sgRNA1/2 and miR-21 sgRNA1 CRISPR-Cas9-edited DU145 cells with a ki67 antibody to evaluate cell proliferation rate. ( F – H ) Labeling MMP9 sgRNA1/2 and miR-21 sgRNA1 CRISPR-Cas9-edited DU145 cells with annexin-5 and 7-AAD to calculate the percentage of cells in the early and late stages of apoptosis and total apoptosis.

Article Snippet: Cells were fixed in 4% paraformaldehyde in PBS for 10 min at room temperature, washed 3 times with PBS and subjected to blocking solution with 3% BSA in PBS for 1 h. The MMP9 (1:150, Boster M00139) and RECK (1:100, Santa Cruz SC373929) antibodies were incubated in PBS containing 1% BSA overnight at 4 °C.

Techniques: Flow Cytometry, Labeling, CRISPR

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer

doi: 10.3390/ijms241914847

Figure Lengend Snippet: Transwell chamber invasion assay with metastatic ( A ) PC-3 and ( B ) DU145 CRISPR-Cas9 MMP9 sgRNA1/2- and miR-21 sgRNA1-edited cell lines.

Article Snippet: Cells were fixed in 4% paraformaldehyde in PBS for 10 min at room temperature, washed 3 times with PBS and subjected to blocking solution with 3% BSA in PBS for 1 h. The MMP9 (1:150, Boster M00139) and RECK (1:100, Santa Cruz SC373929) antibodies were incubated in PBS containing 1% BSA overnight at 4 °C.

Techniques: Invasion Assay, CRISPR