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anti mmp14 mab918  (R&D Systems)


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    R&D Systems anti mmp14 mab918
    Anti Mmp14 Mab918, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+mmp+14/Human+MMP-14%2FMT1-MMP+Antibody/pm41408035-338-13-15
    Average 90 stars, based on 15 article reviews
    anti mmp14 mab918 - by Bioz Stars, 2026-10
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    Article Title: Constitutive Activation of Integrin α9 Augments Self-Directed Hyperplastic and Proinflammatory Properties of Fibroblast-like Synoviocytes of Rheumatoid Arthritis
    Article Snippet: The Abs used in the experiments were as follows: anti-human α9 (rabbit mAb; ab140599; Abcam), anti-human Cadherin-11 (rabbit polyclonal Ab; LS-B2308; LSBio, Seattle, WA), anti–Tn-C (mouse mAb; clone 4F10TT; Immuno-Biological Laboratories, Gunma, Japan), anti–total FAK (rabbit mAb; 3285; Cell Signaling Technology, Danvers, MA), anti-pFAK (pY397) (rabbit mAb; 8556; Cell Signaling Technology), anti–MMP-14 (goat pAb, AF918; R&D Systems), anti-TNFSF11 (goat pAb; AB626; R&D Systems), HRP-conjugated anti-mouse IgG (NA931V; GE Healthcare), and HRP-conjugated anti-rabbit IgG (NA934V; GE Healthcare).



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    anti mmp14 mab918  (R&D Systems)
    90
    R&D Systems anti mmp14 mab918
    Anti Mmp14 Mab918, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+mmp+14/Human+MMP-14%2FMT1-MMP+Antibody/pm41408035-338-13-15
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    anti mt1 mmp antibody  (R&D Systems)
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    R&D Systems anti mt1 mmp antibody
    (A) MDA-MB-231 cells seeded on coverslips, surface labeled with <t>MT1-MMP</t> antibody at 4°C for 1 h Complete media was added, and cells were shifted to 37°C for 30 minutes to allow endocytosis. To remove surface-bound antibodies, an acid wash was given. After washing with PBS, serum-free media was added with or without HGF, and the cells were shifted to 37°C for 10 minutes to allow recycling from the endocytosed pool. Cells were fixed and immunostained for EEA1. The integral intensity of the MT1-MMP and the percentage of MT1-MMP antibody recycling were calculated. N = 3, n= 300; scale bars = 10 µm. The error bar represents means ± SD. Paired t-test, *** P< 0.001, Unpaired t-test, * P < 0.05. (B) MDA-MB-231 cells transfected with pHluorin MT1-MMP were seeded on gelatin-coated glass-bottom dishes. Time-lapse imaging was performed in the presence or absence of HGF stimulation using a Nikon TIRF microscope without any intervals for 300 frames. The flashes of pHluorin MT1-MMP, which represent exocytic events, were calculated using Motiontracking. scale bars = 10 µm. The error bar represents Means ± SEM. Unpaired t-test, ** P < 0.001. N = 4, n= 35. (C) MDA-MB-231 cells were seeded on gelatin-coated glass-bottom dishes and allowed to attach for 3h, followed by incubation with or without HGF for another 3h. Cells were fixed and immunostained for TKS5, MT1-MMP, and F-actin. Images were captured using a Nikon TIRF microscope. TKS5, MT1-MMP, and Actin-positive structures were identified using Motiontracking. The error bar represents Mean ±SEM, Unpaired t-test, * P < 0.01. scale bar: 10µm. N=3, n=120. (D) GFP-MT1-MMP expressing MDA-MB-231 cells were fixed and immunostained for MET. Images were captured using an LSM900 Airyscan super-resolution microscope. Images were processed using the Airyscan Joint deconvolution (jDCV) method. The image was converted to Maximum Intensity projection for representation. The inset shows a magnified view of the region indicated by the box. Scale bar: 10 µm, inset: 2 µm. (E) GFP or GFP-MT1-MMP expressing BT-549 cells were lysed and incubated with GST agarose beads bound to 25µg of GFP binding protein (GBP). Next washes were given to remove unbound proteins and proceeded for immunoblotting with GFP and MET. WCL: whole-cell lysates (F) MDA-MB-231 cells co-expressing GFP-MET and Cherry MT1-MMP were subjected to live-cell TIRF imaging. The inset represents an event at different time points. Scale bar: 10 µm, Inset: 2 µm. (G) BT-549 cells expressing MT1-MMP WT or a catalytic mutant alone or with V5-MET WT or M1250T mutant were seeded on gelatin-coated coverlips for 6h and fixed. Cells were immunostained for V5 and TKS5. Images were captured with the confocal microscope. The number of TKS5 puncta in the MT1-MMP and/or MET expressing cells was calculated and plotted. The error bar represents Mean ±SD, Unpaired t-test. scale bar: 10µm. ** P<0.01, * P < 0.05, N=3, n=150
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    mouse anti mt1 mmp  (R&D Systems)
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    R&D Systems mouse anti mt1 mmp
    (A) MDA-MB-231 cells seeded on coverslips, surface labeled with <t>MT1-MMP</t> antibody at 4°C for 1 h Complete media was added, and cells were shifted to 37°C for 30 minutes to allow endocytosis. To remove surface-bound antibodies, an acid wash was given. After washing with PBS, serum-free media was added with or without HGF, and the cells were shifted to 37°C for 10 minutes to allow recycling from the endocytosed pool. Cells were fixed and immunostained for EEA1. The integral intensity of the MT1-MMP and the percentage of MT1-MMP antibody recycling were calculated. N = 3, n= 300; scale bars = 10 µm. The error bar represents means ± SD. Paired t-test, *** P< 0.001, Unpaired t-test, * P < 0.05. (B) MDA-MB-231 cells transfected with pHluorin MT1-MMP were seeded on gelatin-coated glass-bottom dishes. Time-lapse imaging was performed in the presence or absence of HGF stimulation using a Nikon TIRF microscope without any intervals for 300 frames. The flashes of pHluorin MT1-MMP, which represent exocytic events, were calculated using Motiontracking. scale bars = 10 µm. The error bar represents Means ± SEM. Unpaired t-test, ** P < 0.001. N = 4, n= 35. (C) MDA-MB-231 cells were seeded on gelatin-coated glass-bottom dishes and allowed to attach for 3h, followed by incubation with or without HGF for another 3h. Cells were fixed and immunostained for TKS5, MT1-MMP, and F-actin. Images were captured using a Nikon TIRF microscope. TKS5, MT1-MMP, and Actin-positive structures were identified using Motiontracking. The error bar represents Mean ±SEM, Unpaired t-test, * P < 0.01. scale bar: 10µm. N=3, n=120. (D) GFP-MT1-MMP expressing MDA-MB-231 cells were fixed and immunostained for MET. Images were captured using an LSM900 Airyscan super-resolution microscope. Images were processed using the Airyscan Joint deconvolution (jDCV) method. The image was converted to Maximum Intensity projection for representation. The inset shows a magnified view of the region indicated by the box. Scale bar: 10 µm, inset: 2 µm. (E) GFP or GFP-MT1-MMP expressing BT-549 cells were lysed and incubated with GST agarose beads bound to 25µg of GFP binding protein (GBP). Next washes were given to remove unbound proteins and proceeded for immunoblotting with GFP and MET. WCL: whole-cell lysates (F) MDA-MB-231 cells co-expressing GFP-MET and Cherry MT1-MMP were subjected to live-cell TIRF imaging. The inset represents an event at different time points. Scale bar: 10 µm, Inset: 2 µm. (G) BT-549 cells expressing MT1-MMP WT or a catalytic mutant alone or with V5-MET WT or M1250T mutant were seeded on gelatin-coated coverlips for 6h and fixed. Cells were immunostained for V5 and TKS5. Images were captured with the confocal microscope. The number of TKS5 puncta in the MT1-MMP and/or MET expressing cells was calculated and plotted. The error bar represents Mean ±SD, Unpaired t-test. scale bar: 10µm. ** P<0.01, * P < 0.05, N=3, n=150
    Mouse Anti Mt1 Mmp, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    vii rabbit anti human mt1 mmp mmp 14 cat no 13130 cell signaling technology  (Cell Signaling Technology Inc)
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    Cell Signaling Technology Inc vii rabbit anti human mt1 mmp mmp 14 cat no 13130 cell signaling technology
    (A) MDA-MB-231 cells seeded on coverslips, surface labeled with <t>MT1-MMP</t> antibody at 4°C for 1 h Complete media was added, and cells were shifted to 37°C for 30 minutes to allow endocytosis. To remove surface-bound antibodies, an acid wash was given. After washing with PBS, serum-free media was added with or without HGF, and the cells were shifted to 37°C for 10 minutes to allow recycling from the endocytosed pool. Cells were fixed and immunostained for EEA1. The integral intensity of the MT1-MMP and the percentage of MT1-MMP antibody recycling were calculated. N = 3, n= 300; scale bars = 10 µm. The error bar represents means ± SD. Paired t-test, *** P< 0.001, Unpaired t-test, * P < 0.05. (B) MDA-MB-231 cells transfected with pHluorin MT1-MMP were seeded on gelatin-coated glass-bottom dishes. Time-lapse imaging was performed in the presence or absence of HGF stimulation using a Nikon TIRF microscope without any intervals for 300 frames. The flashes of pHluorin MT1-MMP, which represent exocytic events, were calculated using Motiontracking. scale bars = 10 µm. The error bar represents Means ± SEM. Unpaired t-test, ** P < 0.001. N = 4, n= 35. (C) MDA-MB-231 cells were seeded on gelatin-coated glass-bottom dishes and allowed to attach for 3h, followed by incubation with or without HGF for another 3h. Cells were fixed and immunostained for TKS5, MT1-MMP, and F-actin. Images were captured using a Nikon TIRF microscope. TKS5, MT1-MMP, and Actin-positive structures were identified using Motiontracking. The error bar represents Mean ±SEM, Unpaired t-test, * P < 0.01. scale bar: 10µm. N=3, n=120. (D) GFP-MT1-MMP expressing MDA-MB-231 cells were fixed and immunostained for MET. Images were captured using an LSM900 Airyscan super-resolution microscope. Images were processed using the Airyscan Joint deconvolution (jDCV) method. The image was converted to Maximum Intensity projection for representation. The inset shows a magnified view of the region indicated by the box. Scale bar: 10 µm, inset: 2 µm. (E) GFP or GFP-MT1-MMP expressing BT-549 cells were lysed and incubated with GST agarose beads bound to 25µg of GFP binding protein (GBP). Next washes were given to remove unbound proteins and proceeded for immunoblotting with GFP and MET. WCL: whole-cell lysates (F) MDA-MB-231 cells co-expressing GFP-MET and Cherry MT1-MMP were subjected to live-cell TIRF imaging. The inset represents an event at different time points. Scale bar: 10 µm, Inset: 2 µm. (G) BT-549 cells expressing MT1-MMP WT or a catalytic mutant alone or with V5-MET WT or M1250T mutant were seeded on gelatin-coated coverlips for 6h and fixed. Cells were immunostained for V5 and TKS5. Images were captured with the confocal microscope. The number of TKS5 puncta in the MT1-MMP and/or MET expressing cells was calculated and plotted. The error bar represents Mean ±SD, Unpaired t-test. scale bar: 10µm. ** P<0.01, * P < 0.05, N=3, n=150
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    anti-mmp-14 antibody  (Affinity Biosciences)
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    (A) MDA-MB-231 cells seeded on coverslips, surface labeled with <t>MT1-MMP</t> antibody at 4°C for 1 h Complete media was added, and cells were shifted to 37°C for 30 minutes to allow endocytosis. To remove surface-bound antibodies, an acid wash was given. After washing with PBS, serum-free media was added with or without HGF, and the cells were shifted to 37°C for 10 minutes to allow recycling from the endocytosed pool. Cells were fixed and immunostained for EEA1. The integral intensity of the MT1-MMP and the percentage of MT1-MMP antibody recycling were calculated. N = 3, n= 300; scale bars = 10 µm. The error bar represents means ± SD. Paired t-test, *** P< 0.001, Unpaired t-test, * P < 0.05. (B) MDA-MB-231 cells transfected with pHluorin MT1-MMP were seeded on gelatin-coated glass-bottom dishes. Time-lapse imaging was performed in the presence or absence of HGF stimulation using a Nikon TIRF microscope without any intervals for 300 frames. The flashes of pHluorin MT1-MMP, which represent exocytic events, were calculated using Motiontracking. scale bars = 10 µm. The error bar represents Means ± SEM. Unpaired t-test, ** P < 0.001. N = 4, n= 35. (C) MDA-MB-231 cells were seeded on gelatin-coated glass-bottom dishes and allowed to attach for 3h, followed by incubation with or without HGF for another 3h. Cells were fixed and immunostained for TKS5, MT1-MMP, and F-actin. Images were captured using a Nikon TIRF microscope. TKS5, MT1-MMP, and Actin-positive structures were identified using Motiontracking. The error bar represents Mean ±SEM, Unpaired t-test, * P < 0.01. scale bar: 10µm. N=3, n=120. (D) GFP-MT1-MMP expressing MDA-MB-231 cells were fixed and immunostained for MET. Images were captured using an LSM900 Airyscan super-resolution microscope. Images were processed using the Airyscan Joint deconvolution (jDCV) method. The image was converted to Maximum Intensity projection for representation. The inset shows a magnified view of the region indicated by the box. Scale bar: 10 µm, inset: 2 µm. (E) GFP or GFP-MT1-MMP expressing BT-549 cells were lysed and incubated with GST agarose beads bound to 25µg of GFP binding protein (GBP). Next washes were given to remove unbound proteins and proceeded for immunoblotting with GFP and MET. WCL: whole-cell lysates (F) MDA-MB-231 cells co-expressing GFP-MET and Cherry MT1-MMP were subjected to live-cell TIRF imaging. The inset represents an event at different time points. Scale bar: 10 µm, Inset: 2 µm. (G) BT-549 cells expressing MT1-MMP WT or a catalytic mutant alone or with V5-MET WT or M1250T mutant were seeded on gelatin-coated coverlips for 6h and fixed. Cells were immunostained for V5 and TKS5. Images were captured with the confocal microscope. The number of TKS5 puncta in the MT1-MMP and/or MET expressing cells was calculated and plotted. The error bar represents Mean ±SD, Unpaired t-test. scale bar: 10µm. ** P<0.01, * P < 0.05, N=3, n=150
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    Single-cell RNA-Seq results of nasopharyngeal/pharyngeal swabs, bronchial brushings and bronchial lavages were generated from patients with COVID-19 ( n = 19) and healthy controls ( n = 5). Data are extracted from Chua et al. . ( A ) Both ANPEP (codes for CD13) and <t>MMP14</t> are expressed on various epithelial cells and macrophages. ANPEP is also expressed on neutrophils while MMP14 is expressed on mast cells. BDKRB1 (codes for B1R) is expressed on some epithelial cells with the highest expression on secretory cells and ciliated cells. F2RL3 (codes for PAR4) was barely detected in this dataset but seemed to be expressed by epithelial cells. Cell abbreviations are defined in . ( B ) Cellular ANPEP expression in nasopharyngeal cells is elevated in patients with COVID-19 (moderate COVID-19 n = 8, severe COVID-19 n = 11) compared with healthy controls ( n = 5). ( C ) Using the median expression levels of ANPEP in patients, patients with COVID-19 were divided into 2 populations: ANPEP -high and ANPEP -low. Differentially expressed genes in neutrophils from these 2 groups were shown in the volcano plot ( P < 1 × 10 –20 , |log 2 (fold-change)| < 0.25). ( D ) Pathway analysis of the differentially expressed genes in ANPEP -high and ANPEP -low neutrophils showed pathways related to neutrophil degranulation, leukocyte activation, and inflammation. ( E ) The neutrophils from ANPEP -high patients with COVID-19 ( n = 10) showed a gene signature of immature-like neutrophils characterized by the overexpression of genes coding for several granule-content proteins (healthy controls n = 5, ANPEP -low n = 9). ( F ) The score for neutrophil immaturity was higher in critically ill patients with COVID-19 ( n = 11) compared with moderate patients (left, n = 8). The median neutrophil-immaturity score of neutrophils in ANPEP -high patients ( n = 10) was lower than that in ANPEP -low patients (right, n = 9). ( G ) The neutrophil-immaturity signature was most prominent in ANPEP -high patients ( n = 10) as developing neutrophils versus mature neutrophils from the peripheral blood were examined (healthy controls n = 5, ANPEP -low n = 9). Results are expressed as mean ± SD. **** P < 0.0001. Significance was determined by 1-way ANOVA ( B and G ) and Mann-Whitney test ( F ).
    Mmp14, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti mmp 14 antibodies  (Santa Cruz Biotechnology)
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    Santa Cruz Biotechnology anti mmp 14 antibodies
    Single-cell RNA-Seq results of nasopharyngeal/pharyngeal swabs, bronchial brushings and bronchial lavages were generated from patients with COVID-19 ( n = 19) and healthy controls ( n = 5). Data are extracted from Chua et al. . ( A ) Both ANPEP (codes for CD13) and <t>MMP14</t> are expressed on various epithelial cells and macrophages. ANPEP is also expressed on neutrophils while MMP14 is expressed on mast cells. BDKRB1 (codes for B1R) is expressed on some epithelial cells with the highest expression on secretory cells and ciliated cells. F2RL3 (codes for PAR4) was barely detected in this dataset but seemed to be expressed by epithelial cells. Cell abbreviations are defined in . ( B ) Cellular ANPEP expression in nasopharyngeal cells is elevated in patients with COVID-19 (moderate COVID-19 n = 8, severe COVID-19 n = 11) compared with healthy controls ( n = 5). ( C ) Using the median expression levels of ANPEP in patients, patients with COVID-19 were divided into 2 populations: ANPEP -high and ANPEP -low. Differentially expressed genes in neutrophils from these 2 groups were shown in the volcano plot ( P < 1 × 10 –20 , |log 2 (fold-change)| < 0.25). ( D ) Pathway analysis of the differentially expressed genes in ANPEP -high and ANPEP -low neutrophils showed pathways related to neutrophil degranulation, leukocyte activation, and inflammation. ( E ) The neutrophils from ANPEP -high patients with COVID-19 ( n = 10) showed a gene signature of immature-like neutrophils characterized by the overexpression of genes coding for several granule-content proteins (healthy controls n = 5, ANPEP -low n = 9). ( F ) The score for neutrophil immaturity was higher in critically ill patients with COVID-19 ( n = 11) compared with moderate patients (left, n = 8). The median neutrophil-immaturity score of neutrophils in ANPEP -high patients ( n = 10) was lower than that in ANPEP -low patients (right, n = 9). ( G ) The neutrophil-immaturity signature was most prominent in ANPEP -high patients ( n = 10) as developing neutrophils versus mature neutrophils from the peripheral blood were examined (healthy controls n = 5, ANPEP -low n = 9). Results are expressed as mean ± SD. **** P < 0.0001. Significance was determined by 1-way ANOVA ( B and G ) and Mann-Whitney test ( F ).
    Anti Mmp 14 Antibodies, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mmp 14  (Cell Signaling Technology Inc)
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    Single-cell RNA-Seq results of nasopharyngeal/pharyngeal swabs, bronchial brushings and bronchial lavages were generated from patients with COVID-19 ( n = 19) and healthy controls ( n = 5). Data are extracted from Chua et al. . ( A ) Both ANPEP (codes for CD13) and <t>MMP14</t> are expressed on various epithelial cells and macrophages. ANPEP is also expressed on neutrophils while MMP14 is expressed on mast cells. BDKRB1 (codes for B1R) is expressed on some epithelial cells with the highest expression on secretory cells and ciliated cells. F2RL3 (codes for PAR4) was barely detected in this dataset but seemed to be expressed by epithelial cells. Cell abbreviations are defined in . ( B ) Cellular ANPEP expression in nasopharyngeal cells is elevated in patients with COVID-19 (moderate COVID-19 n = 8, severe COVID-19 n = 11) compared with healthy controls ( n = 5). ( C ) Using the median expression levels of ANPEP in patients, patients with COVID-19 were divided into 2 populations: ANPEP -high and ANPEP -low. Differentially expressed genes in neutrophils from these 2 groups were shown in the volcano plot ( P < 1 × 10 –20 , |log 2 (fold-change)| < 0.25). ( D ) Pathway analysis of the differentially expressed genes in ANPEP -high and ANPEP -low neutrophils showed pathways related to neutrophil degranulation, leukocyte activation, and inflammation. ( E ) The neutrophils from ANPEP -high patients with COVID-19 ( n = 10) showed a gene signature of immature-like neutrophils characterized by the overexpression of genes coding for several granule-content proteins (healthy controls n = 5, ANPEP -low n = 9). ( F ) The score for neutrophil immaturity was higher in critically ill patients with COVID-19 ( n = 11) compared with moderate patients (left, n = 8). The median neutrophil-immaturity score of neutrophils in ANPEP -high patients ( n = 10) was lower than that in ANPEP -low patients (right, n = 9). ( G ) The neutrophil-immaturity signature was most prominent in ANPEP -high patients ( n = 10) as developing neutrophils versus mature neutrophils from the peripheral blood were examined (healthy controls n = 5, ANPEP -low n = 9). Results are expressed as mean ± SD. **** P < 0.0001. Significance was determined by 1-way ANOVA ( B and G ) and Mann-Whitney test ( F ).
    Mmp 14, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    (A) MDA-MB-231 cells seeded on coverslips, surface labeled with MT1-MMP antibody at 4°C for 1 h Complete media was added, and cells were shifted to 37°C for 30 minutes to allow endocytosis. To remove surface-bound antibodies, an acid wash was given. After washing with PBS, serum-free media was added with or without HGF, and the cells were shifted to 37°C for 10 minutes to allow recycling from the endocytosed pool. Cells were fixed and immunostained for EEA1. The integral intensity of the MT1-MMP and the percentage of MT1-MMP antibody recycling were calculated. N = 3, n= 300; scale bars = 10 µm. The error bar represents means ± SD. Paired t-test, *** P< 0.001, Unpaired t-test, * P < 0.05. (B) MDA-MB-231 cells transfected with pHluorin MT1-MMP were seeded on gelatin-coated glass-bottom dishes. Time-lapse imaging was performed in the presence or absence of HGF stimulation using a Nikon TIRF microscope without any intervals for 300 frames. The flashes of pHluorin MT1-MMP, which represent exocytic events, were calculated using Motiontracking. scale bars = 10 µm. The error bar represents Means ± SEM. Unpaired t-test, ** P < 0.001. N = 4, n= 35. (C) MDA-MB-231 cells were seeded on gelatin-coated glass-bottom dishes and allowed to attach for 3h, followed by incubation with or without HGF for another 3h. Cells were fixed and immunostained for TKS5, MT1-MMP, and F-actin. Images were captured using a Nikon TIRF microscope. TKS5, MT1-MMP, and Actin-positive structures were identified using Motiontracking. The error bar represents Mean ±SEM, Unpaired t-test, * P < 0.01. scale bar: 10µm. N=3, n=120. (D) GFP-MT1-MMP expressing MDA-MB-231 cells were fixed and immunostained for MET. Images were captured using an LSM900 Airyscan super-resolution microscope. Images were processed using the Airyscan Joint deconvolution (jDCV) method. The image was converted to Maximum Intensity projection for representation. The inset shows a magnified view of the region indicated by the box. Scale bar: 10 µm, inset: 2 µm. (E) GFP or GFP-MT1-MMP expressing BT-549 cells were lysed and incubated with GST agarose beads bound to 25µg of GFP binding protein (GBP). Next washes were given to remove unbound proteins and proceeded for immunoblotting with GFP and MET. WCL: whole-cell lysates (F) MDA-MB-231 cells co-expressing GFP-MET and Cherry MT1-MMP were subjected to live-cell TIRF imaging. The inset represents an event at different time points. Scale bar: 10 µm, Inset: 2 µm. (G) BT-549 cells expressing MT1-MMP WT or a catalytic mutant alone or with V5-MET WT or M1250T mutant were seeded on gelatin-coated coverlips for 6h and fixed. Cells were immunostained for V5 and TKS5. Images were captured with the confocal microscope. The number of TKS5 puncta in the MT1-MMP and/or MET expressing cells was calculated and plotted. The error bar represents Mean ±SD, Unpaired t-test. scale bar: 10µm. ** P<0.01, * P < 0.05, N=3, n=150

    Journal: bioRxiv

    Article Title: RAB14-dependent tubulovesicular recycling directs MET to invadopodia promoting TNBC cell invasion

    doi: 10.1101/2025.09.23.677683

    Figure Lengend Snippet: (A) MDA-MB-231 cells seeded on coverslips, surface labeled with MT1-MMP antibody at 4°C for 1 h Complete media was added, and cells were shifted to 37°C for 30 minutes to allow endocytosis. To remove surface-bound antibodies, an acid wash was given. After washing with PBS, serum-free media was added with or without HGF, and the cells were shifted to 37°C for 10 minutes to allow recycling from the endocytosed pool. Cells were fixed and immunostained for EEA1. The integral intensity of the MT1-MMP and the percentage of MT1-MMP antibody recycling were calculated. N = 3, n= 300; scale bars = 10 µm. The error bar represents means ± SD. Paired t-test, *** P< 0.001, Unpaired t-test, * P < 0.05. (B) MDA-MB-231 cells transfected with pHluorin MT1-MMP were seeded on gelatin-coated glass-bottom dishes. Time-lapse imaging was performed in the presence or absence of HGF stimulation using a Nikon TIRF microscope without any intervals for 300 frames. The flashes of pHluorin MT1-MMP, which represent exocytic events, were calculated using Motiontracking. scale bars = 10 µm. The error bar represents Means ± SEM. Unpaired t-test, ** P < 0.001. N = 4, n= 35. (C) MDA-MB-231 cells were seeded on gelatin-coated glass-bottom dishes and allowed to attach for 3h, followed by incubation with or without HGF for another 3h. Cells were fixed and immunostained for TKS5, MT1-MMP, and F-actin. Images were captured using a Nikon TIRF microscope. TKS5, MT1-MMP, and Actin-positive structures were identified using Motiontracking. The error bar represents Mean ±SEM, Unpaired t-test, * P < 0.01. scale bar: 10µm. N=3, n=120. (D) GFP-MT1-MMP expressing MDA-MB-231 cells were fixed and immunostained for MET. Images were captured using an LSM900 Airyscan super-resolution microscope. Images were processed using the Airyscan Joint deconvolution (jDCV) method. The image was converted to Maximum Intensity projection for representation. The inset shows a magnified view of the region indicated by the box. Scale bar: 10 µm, inset: 2 µm. (E) GFP or GFP-MT1-MMP expressing BT-549 cells were lysed and incubated with GST agarose beads bound to 25µg of GFP binding protein (GBP). Next washes were given to remove unbound proteins and proceeded for immunoblotting with GFP and MET. WCL: whole-cell lysates (F) MDA-MB-231 cells co-expressing GFP-MET and Cherry MT1-MMP were subjected to live-cell TIRF imaging. The inset represents an event at different time points. Scale bar: 10 µm, Inset: 2 µm. (G) BT-549 cells expressing MT1-MMP WT or a catalytic mutant alone or with V5-MET WT or M1250T mutant were seeded on gelatin-coated coverlips for 6h and fixed. Cells were immunostained for V5 and TKS5. Images were captured with the confocal microscope. The number of TKS5 puncta in the MT1-MMP and/or MET expressing cells was calculated and plotted. The error bar represents Mean ±SD, Unpaired t-test. scale bar: 10µm. ** P<0.01, * P < 0.05, N=3, n=150

    Article Snippet: MDA-MB-231 cells were trypsinized and counted, and 50,000 cells per well were seeded on the gelatin-coated coverslips for 24 h. After the cells had properly adhered and regained their morphology, antibody uptake was performed using anti-MT1-MMP antibody (R&D Systems, MAB9181-SP).

    Techniques: Labeling, Transfection, Imaging, Microscopy, Incubation, Expressing, Super-Resolution Microscopy, Binding Assay, Western Blot, Mutagenesis

    HGF stimulation triggers invadopodia formation and ECM degradation by promoting invadopodia formation and MT1-MMP recycling. HGF also increases the recruitment of MET to invadopodia by stimulating the trafficking of the RTK in the RAB4 and RAB14-dependent trafficking pathway. HGF stimulation increases the phosphorylation and colocalization of RCP with MET-containing RAB14 vesicles. RCP and MET are present on the endosomal budding site of RAB14, which further elongate and form tubules. KIF16B gets recruited to these endosomes through RAB14 and drives the endosomal tubulation for efficient cargo trafficking. MT1-MMP interacts with MET, presumably promoting their co-trafficking to invadopodia.

    Journal: bioRxiv

    Article Title: RAB14-dependent tubulovesicular recycling directs MET to invadopodia promoting TNBC cell invasion

    doi: 10.1101/2025.09.23.677683

    Figure Lengend Snippet: HGF stimulation triggers invadopodia formation and ECM degradation by promoting invadopodia formation and MT1-MMP recycling. HGF also increases the recruitment of MET to invadopodia by stimulating the trafficking of the RTK in the RAB4 and RAB14-dependent trafficking pathway. HGF stimulation increases the phosphorylation and colocalization of RCP with MET-containing RAB14 vesicles. RCP and MET are present on the endosomal budding site of RAB14, which further elongate and form tubules. KIF16B gets recruited to these endosomes through RAB14 and drives the endosomal tubulation for efficient cargo trafficking. MT1-MMP interacts with MET, presumably promoting their co-trafficking to invadopodia.

    Article Snippet: MDA-MB-231 cells were trypsinized and counted, and 50,000 cells per well were seeded on the gelatin-coated coverslips for 24 h. After the cells had properly adhered and regained their morphology, antibody uptake was performed using anti-MT1-MMP antibody (R&D Systems, MAB9181-SP).

    Techniques: Phospho-proteomics

    Single-cell RNA-Seq results of nasopharyngeal/pharyngeal swabs, bronchial brushings and bronchial lavages were generated from patients with COVID-19 ( n = 19) and healthy controls ( n = 5). Data are extracted from Chua et al. . ( A ) Both ANPEP (codes for CD13) and MMP14 are expressed on various epithelial cells and macrophages. ANPEP is also expressed on neutrophils while MMP14 is expressed on mast cells. BDKRB1 (codes for B1R) is expressed on some epithelial cells with the highest expression on secretory cells and ciliated cells. F2RL3 (codes for PAR4) was barely detected in this dataset but seemed to be expressed by epithelial cells. Cell abbreviations are defined in . ( B ) Cellular ANPEP expression in nasopharyngeal cells is elevated in patients with COVID-19 (moderate COVID-19 n = 8, severe COVID-19 n = 11) compared with healthy controls ( n = 5). ( C ) Using the median expression levels of ANPEP in patients, patients with COVID-19 were divided into 2 populations: ANPEP -high and ANPEP -low. Differentially expressed genes in neutrophils from these 2 groups were shown in the volcano plot ( P < 1 × 10 –20 , |log 2 (fold-change)| < 0.25). ( D ) Pathway analysis of the differentially expressed genes in ANPEP -high and ANPEP -low neutrophils showed pathways related to neutrophil degranulation, leukocyte activation, and inflammation. ( E ) The neutrophils from ANPEP -high patients with COVID-19 ( n = 10) showed a gene signature of immature-like neutrophils characterized by the overexpression of genes coding for several granule-content proteins (healthy controls n = 5, ANPEP -low n = 9). ( F ) The score for neutrophil immaturity was higher in critically ill patients with COVID-19 ( n = 11) compared with moderate patients (left, n = 8). The median neutrophil-immaturity score of neutrophils in ANPEP -high patients ( n = 10) was lower than that in ANPEP -low patients (right, n = 9). ( G ) The neutrophil-immaturity signature was most prominent in ANPEP -high patients ( n = 10) as developing neutrophils versus mature neutrophils from the peripheral blood were examined (healthy controls n = 5, ANPEP -low n = 9). Results are expressed as mean ± SD. **** P < 0.0001. Significance was determined by 1-way ANOVA ( B and G ) and Mann-Whitney test ( F ).

    Journal: JCI Insight

    Article Title: Soluble CD13 is a potential mediator of neutrophil-induced thrombogenic inflammation in SARS-CoV-2 infection

    doi: 10.1172/jci.insight.184975

    Figure Lengend Snippet: Single-cell RNA-Seq results of nasopharyngeal/pharyngeal swabs, bronchial brushings and bronchial lavages were generated from patients with COVID-19 ( n = 19) and healthy controls ( n = 5). Data are extracted from Chua et al. . ( A ) Both ANPEP (codes for CD13) and MMP14 are expressed on various epithelial cells and macrophages. ANPEP is also expressed on neutrophils while MMP14 is expressed on mast cells. BDKRB1 (codes for B1R) is expressed on some epithelial cells with the highest expression on secretory cells and ciliated cells. F2RL3 (codes for PAR4) was barely detected in this dataset but seemed to be expressed by epithelial cells. Cell abbreviations are defined in . ( B ) Cellular ANPEP expression in nasopharyngeal cells is elevated in patients with COVID-19 (moderate COVID-19 n = 8, severe COVID-19 n = 11) compared with healthy controls ( n = 5). ( C ) Using the median expression levels of ANPEP in patients, patients with COVID-19 were divided into 2 populations: ANPEP -high and ANPEP -low. Differentially expressed genes in neutrophils from these 2 groups were shown in the volcano plot ( P < 1 × 10 –20 , |log 2 (fold-change)| < 0.25). ( D ) Pathway analysis of the differentially expressed genes in ANPEP -high and ANPEP -low neutrophils showed pathways related to neutrophil degranulation, leukocyte activation, and inflammation. ( E ) The neutrophils from ANPEP -high patients with COVID-19 ( n = 10) showed a gene signature of immature-like neutrophils characterized by the overexpression of genes coding for several granule-content proteins (healthy controls n = 5, ANPEP -low n = 9). ( F ) The score for neutrophil immaturity was higher in critically ill patients with COVID-19 ( n = 11) compared with moderate patients (left, n = 8). The median neutrophil-immaturity score of neutrophils in ANPEP -high patients ( n = 10) was lower than that in ANPEP -low patients (right, n = 9). ( G ) The neutrophil-immaturity signature was most prominent in ANPEP -high patients ( n = 10) as developing neutrophils versus mature neutrophils from the peripheral blood were examined (healthy controls n = 5, ANPEP -low n = 9). Results are expressed as mean ± SD. **** P < 0.0001. Significance was determined by 1-way ANOVA ( B and G ) and Mann-Whitney test ( F ).

    Article Snippet: White blood cells in the upper phase were washed with PBS containing 2 mM EDTA and 2% FBS and stained with fluorochrome-conjugated antibodies against human CD3 (PE-Cy7, BioLegend 300420), CD13 (PE, BioLegend 301703), CD14 (APC-Cy7, BioLegend 301820) CD16 (PerCPCy5.5, Tonbo 615-1619-T025), CD19 (BV510, BioLegend 302241), CD45 (APC-Cy7, Tonbo 25-0459-T100), CD56 (PE, BioLegend 362508), CD10 (Super Bright 702, Invitrogen 67-010-642), PAR4 (FITC, Alomone Labs APR-034-F), B1R (APC, LS Bio LS-C275585), MMP14 (APC, R&D Systems, Bio-Techne; FAB9181A), and CD66b (Pacific Blue, BioLegend 305111).

    Techniques: RNA Sequencing, Generated, Expressing, Activation Assay, Over Expression, MANN-WHITNEY

    ( A ) Gating scheme of flow cytometry analysis on monocytes and neutrophils. ( B ) CD13, B1R, and PAR4 were expressed on monocytes and neutrophils, while MMP14 showed minimal expression on these cells ( n = 3). ( C ) Histograms showing the changes in CD13, PAR4, and B1R expression in neutrophils and monocytes with or without IL-1β, TNF-α, or IL-6 stimulation. ( D ) Quantification of relative expression of CD13, PAR4, and B1R on neutrophils and monocytes after stimulation with IL-1β, TNF-α, or IL-6 compared with unstimulated control from 3 healthy donors. Results are expressed as mean ± SD. * P < 0.05, ** P < 0.01, **** P < 0.0001. Significance was determined by 1-way ANOVA. FMO, fluorescence minus 1 (gating control); NT, not treated.

    Journal: JCI Insight

    Article Title: Soluble CD13 is a potential mediator of neutrophil-induced thrombogenic inflammation in SARS-CoV-2 infection

    doi: 10.1172/jci.insight.184975

    Figure Lengend Snippet: ( A ) Gating scheme of flow cytometry analysis on monocytes and neutrophils. ( B ) CD13, B1R, and PAR4 were expressed on monocytes and neutrophils, while MMP14 showed minimal expression on these cells ( n = 3). ( C ) Histograms showing the changes in CD13, PAR4, and B1R expression in neutrophils and monocytes with or without IL-1β, TNF-α, or IL-6 stimulation. ( D ) Quantification of relative expression of CD13, PAR4, and B1R on neutrophils and monocytes after stimulation with IL-1β, TNF-α, or IL-6 compared with unstimulated control from 3 healthy donors. Results are expressed as mean ± SD. * P < 0.05, ** P < 0.01, **** P < 0.0001. Significance was determined by 1-way ANOVA. FMO, fluorescence minus 1 (gating control); NT, not treated.

    Article Snippet: White blood cells in the upper phase were washed with PBS containing 2 mM EDTA and 2% FBS and stained with fluorochrome-conjugated antibodies against human CD3 (PE-Cy7, BioLegend 300420), CD13 (PE, BioLegend 301703), CD14 (APC-Cy7, BioLegend 301820) CD16 (PerCPCy5.5, Tonbo 615-1619-T025), CD19 (BV510, BioLegend 302241), CD45 (APC-Cy7, Tonbo 25-0459-T100), CD56 (PE, BioLegend 362508), CD10 (Super Bright 702, Invitrogen 67-010-642), PAR4 (FITC, Alomone Labs APR-034-F), B1R (APC, LS Bio LS-C275585), MMP14 (APC, R&D Systems, Bio-Techne; FAB9181A), and CD66b (Pacific Blue, BioLegend 305111).

    Techniques: Flow Cytometry, Expressing, Control, Fluorescence