Journal: Science Advances

Article Title: An injury-responsive mmp14b enhancer is required for heart regeneration

doi: 10.1126/sciadv.adh5313

Figure Lengend Snippet: ( A ) Schematic of the experimental design for neonatal mouse heart injury and MMP-14 inhibitor treatment. Myocardial injury by coronary artery ligation was conducted on P1, mice were simultaneously treated with a single intraperitoneal injection of 20 μM NSC405020 or DMSO (vehicle control), and hearts were collected on P4 for analysis by Western blot (WB). ( B ) Western blots for Agrin and β-actin from total heart tissue 3 dpi. ( C ) Quantification of the 260-kD isoform of Agrin protein normalized to β-actin and expressed in AU in neonatal mouse hearts following myocardial injury (3 dpi) and treatment with either DMSO or NSC405020. Statistical significance was determined using Student’s t test. * P < 0.05.

Article Snippet: For inhibitor studies, following amputation, zebrafish were divided into 500-ml tanks, the MT1-MMP inhibitor NSC405020 (Sigma-Aldrich) was added in DMSO at a final inhibitor concentration of 20 μM, and caudal fin regeneration in the presence of inhibitor, compared to the addition of 0.1% DMSO alone, was assessed at the indicated time points.

Techniques: Ligation, Injection, Western Blot

Journal: Molecular Oncology

Article Title: MMP14 expression and collagen remodelling support uterine leiomyosarcoma aggressiveness

doi: 10.1002/1878-0261.13440

Figure Lengend Snippet: uLMS cells highly express MMP14 and depend on its activity for proliferation in collagenous environments. (A–C), Relative average expression of fibrillar collagen (fCOL) (A), collagen biosynthesis and crosslinking (COLbio) (B), and collagen‐cleaving matrix metalloproteinases (MMPs) (C), in normal myometrium (MM; n = 31), uterine leiomyoma (LM; n = 28), and uterine leiomyosarcoma (uLMS; n = 17). One‐way ANOVA with post hoc Tukey's test. (D) Relative average expression of MMPs in uLMS and other uterine sarcomas included in the cohort (other Sarc; n = 17). Two‐tailed Student's t ‐test. (E) Representative examples of MMP14 protein staining in LM ( n = 12) and uLMS ( n = 9) tissues. Scale bar indicates 100 μm. (F) Quantification of the percentage of the area of the tissue positive for MMP14 in LM ( n = 12) and uLMS ( n = 9) tissues, indicating higher MMP14 protein expression in uLMS. Each datapoint represents one patient; horizontal bars indicate the average per tissue type. Two‐tailed Student's t ‐test. (G) Association between high expression of fCOL, COLbio and MMP genes and uLMS patient prognosis of the TCGA cohort ( n = 24). Patients groups were generated by the best cut‐off method, favourable/unfavourable survival was defined as significant changes of median survival ( P < 0.05). (H, I) Metascape pathway analysis of the genes upregulated (H) and downregulated (I) in uLMS tumours with low fraction of high‐density matrix (HDMl ow ; n = 10) compared with HDM high ( n = 8) uLMS tumours. (J) Representative images of SKUT1 cells in 3D collagen gels 4 days after MMP14 knockdown ( n = 3 independent experiments). Scale bar indicates 50 μm. (K) Proliferative (Ki67+) and apoptotic (Cleaved‐caspase 3+) cell quantification from (J) ( n = 3 independent experiments). One‐way ANOVA with post hoc Tukey's test. (L) Quantification of the response of patient‐derived uLMS and low‐grade endometrial stromal sarcoma (LG‐ESS) cells to NSC405020. Each data point indicated the average of 3 technical replicates. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cells were treated with GM6001 (pan‐MMP; Calbiochem, San Diego, CA, USA), NSC405020 (MMP14‐specific; Selleckchem, Houston, TX, USA) or verteporfin (YAP; Sigma) for the indicated times and concentrations.

Techniques: Activity Assay, Expressing, Two Tailed Test, Staining, Generated, Knockdown, Derivative Assay

Journal: Nature immunology

Article Title: Fungal sensing by dectin-1 directs the non-pathogenic polarization of T H 17 cells through balanced type I IFN responses in human DCs.

doi: 10.1038/s41590-022-01348-2

Figure Lengend Snippet: Fig. 7 | Dectin-1-induced release of active TGF-β requires MMP14 activity. a, SEAP assay on supernatant of HEK-Blue TGF-β reporter cells for quantification of active TGF-β in the supernatant of unstimulated DCs or after stimulation of DCs with curdlan or C. albicans, in the presence of MMP14 inhibitor NSC405020 at 24 h (n = 3). b–e, FRET assay of extracellular MMP14 activity (RFU) in unstimulated DCs or after stimulation of DCs with curdlan or C. albicans, in the presence of NSC405020 (b), blocking IFN-α/βR antibodies (c), a concentration

Article Snippet: DCs were preincubated for 2 h with MMP14 inhibitor NSC405020 (100 μM; Tocris) or blocking antibodies, anti-dectin-1 (20 μg ml−1; clone 259931, MAB1859, R&D Systems), anti-IFN-α/βR2 (20 μg ml−1; clone MMHAR-2, PBL Assay Science), anti-αvβ1 (10 μg ml−1; clone P5D2, MAB17781, R&D Systems), anti-αvβ3 (10 μg ml−1; clone 23C6, MAB3050, R&D Systems), anti-αvβ5 (10 μg ml−1; clone P5H9, MAB2528, R&D Systems), anti-αvβ6 (10 μg ml−1; clone 10D5, ab77906, Abcam), anti-αvβ8 (10 μg ml−1; kind gift from S.L.

Techniques: Activity Assay, SEAP Assay, Blocking Assay, Concentration Assay

Journal: Biosensors

Article Title: Detecting the PEX Like Domain of Matrix Metalloproteinase-14 (MMP-14) with Therapeutic Conjugated CNTs

doi: 10.3390/bios12100884

Figure Lengend Snippet: ( A ) Schematic representation of NSC405020 molecule grafted onto CNTs surface followed by ( B ) SEM images for pristine, acylated, and inhibitor loaded CNTs (low and high magnification).

Article Snippet: 3,4-Dichloro-N-(pentan-2-yl) benzamide, (MMP-14 Inhibitor- NSC405020 (AmBeed, Arlington Heights, IL, USA)) was grafted onto acylated CNTs.

Techniques:

Journal: Biosensors

Article Title: Detecting the PEX Like Domain of Matrix Metalloproteinase-14 (MMP-14) with Therapeutic Conjugated CNTs

doi: 10.3390/bios12100884

Figure Lengend Snippet: ( A ) Nyquist plots of inhibitor loaded CNTs before and after interaction with different concentrations of MPP-14 for 10 minutes in PBS (pH 7.40) containing 10 mmol.mL −1 of K 3 [Fe (CN) 6 ] − and 10 mmol.mL −1 of NaCl. ( B ) Linear fit of EIS response for different concentrations of MMP-14, presenting LOD of 7.5 ng⋅mL −1 . Significant difference between measurements was obtained at p -value < 0.05 (n = 2). Protein concentration varying from 10 ng.mL −1 to 100 ng.mL −1 , applied potential of +0.20 V, from 50 kHz to 500 Hz, amplitude 50 mV.

Article Snippet: 3,4-Dichloro-N-(pentan-2-yl) benzamide, (MMP-14 Inhibitor- NSC405020 (AmBeed, Arlington Heights, IL, USA)) was grafted onto acylated CNTs.

Techniques: Protein Concentration

Journal: Biosensors

Article Title: Detecting the PEX Like Domain of Matrix Metalloproteinase-14 (MMP-14) with Therapeutic Conjugated CNTs

doi: 10.3390/bios12100884

Figure Lengend Snippet: ( A ) The inhibitor/protein interaction between small molecule NSC 405020 and the binding pocket of MMP-14 and ( B ) Schematic of the MMP-14 detection through the binding mechanism of protein and inhibitory small molecule as the recognition element.

Article Snippet: 3,4-Dichloro-N-(pentan-2-yl) benzamide, (MMP-14 Inhibitor- NSC405020 (AmBeed, Arlington Heights, IL, USA)) was grafted onto acylated CNTs.

Techniques: Binding Assay

Journal: Biosensors

Article Title: Detecting the PEX Like Domain of Matrix Metalloproteinase-14 (MMP-14) with Therapeutic Conjugated CNTs

doi: 10.3390/bios12100884

Figure Lengend Snippet: EIS response for the inhibitor loaded CNT after individual interaction with 50 ng⋅mL −1 of MMP-1 and MMP-14 in PBS (pH 7.40) containing 10 mmol⋅mL −1 of K 4 [Fe (CN) 6 ] − 10 mmol⋅mL −1 of K 3 [Fe (CN) 6 ] − and 10 mmol⋅mL −1 of NaCl. The inhibitor loaded CNT showed specificity to MMP-14 (applied potential of +0.20 V, from 50 kHz to 500 Hz, amplitude 50 mV).

Article Snippet: 3,4-Dichloro-N-(pentan-2-yl) benzamide, (MMP-14 Inhibitor- NSC405020 (AmBeed, Arlington Heights, IL, USA)) was grafted onto acylated CNTs.

Techniques: