collagenase type i  (Worthington Biochemical)


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    Name:
    Collagenase Type 1
    Description:
    The original balance of enzymatic activities Each lot assayed for collagenase caseinase clostripain and tryptic activities Suggested for epithelial liver lung and adrenal primary cell isolations A dialyzed lyophilized powder
    Catalog Number:
    LS004194
    Price:
    35
    Source:
    Clostridium histolyticum
    Size:
    100 mg
    Cas Number:
    9001.12.1
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    Structured Review

    Worthington Biochemical collagenase type i
    SMC migration was reduced in the absence of DDR1. SMCs from DDR1+/+ mice (filled bars) or DDR1–/– mice (open bars) were stimulated to migrate for 4 hours in chemotaxis chambers with 200 nM <t>type</t> I collagen (left) or type VIII collagen (right) added to <t>DMEM</t> in the bottom of the chamber. Migration was quantified by staining and counting the number of cells that migrated to the bottom of the filter. Experiments were done in triplicate and repeated three times. A Values from DDR1–/– SMCs were significantly less than values from wild-type SMCs.
    The original balance of enzymatic activities Each lot assayed for collagenase caseinase clostripain and tryptic activities Suggested for epithelial liver lung and adrenal primary cell isolations A dialyzed lyophilized powder
    https://www.bioz.com/result/collagenase type i/product/Worthington Biochemical
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    collagenase type i - by Bioz Stars, 2021-04
    99/100 stars

    Images

    1) Product Images from "The discoidin domain receptor tyrosine kinase DDR1 in arterial wound repair"

    Article Title: The discoidin domain receptor tyrosine kinase DDR1 in arterial wound repair

    Journal: Journal of Clinical Investigation

    doi:

    SMC migration was reduced in the absence of DDR1. SMCs from DDR1+/+ mice (filled bars) or DDR1–/– mice (open bars) were stimulated to migrate for 4 hours in chemotaxis chambers with 200 nM type I collagen (left) or type VIII collagen (right) added to DMEM in the bottom of the chamber. Migration was quantified by staining and counting the number of cells that migrated to the bottom of the filter. Experiments were done in triplicate and repeated three times. A Values from DDR1–/– SMCs were significantly less than values from wild-type SMCs.
    Figure Legend Snippet: SMC migration was reduced in the absence of DDR1. SMCs from DDR1+/+ mice (filled bars) or DDR1–/– mice (open bars) were stimulated to migrate for 4 hours in chemotaxis chambers with 200 nM type I collagen (left) or type VIII collagen (right) added to DMEM in the bottom of the chamber. Migration was quantified by staining and counting the number of cells that migrated to the bottom of the filter. Experiments were done in triplicate and repeated three times. A Values from DDR1–/– SMCs were significantly less than values from wild-type SMCs.

    Techniques Used: Migration, Mouse Assay, Chemotaxis Assay, Staining

    Cell proliferation on collagen was less in DDR1–/– SMCs compared with DDR1+/+ SMCs. ( a ) SMCs from DDR1+/+ mice (filled bars) or DDR1–/– mice (open bars) were stimulated to grow with 10% FCS and incubated with [ 3 H]thymidine on wells coated with 100 nM type I collagen (left) or type VIII collagen (right). A Value from DDR1–/– SMCs was significantly less than value from wild-type SMCs. ( b ) Number of SMCs per well after plating on 100 nM type I collagen and growing in DMEM containing 10% FCS for 1–4 days. Squares, DDR1+/+ SMCs; circles, DDR1–/– SMCs. Experiments were done in triplicate and repeated three times. A Value from DDR1–/– SMCs was significantly less than value from wild-type SMCs.
    Figure Legend Snippet: Cell proliferation on collagen was less in DDR1–/– SMCs compared with DDR1+/+ SMCs. ( a ) SMCs from DDR1+/+ mice (filled bars) or DDR1–/– mice (open bars) were stimulated to grow with 10% FCS and incubated with [ 3 H]thymidine on wells coated with 100 nM type I collagen (left) or type VIII collagen (right). A Value from DDR1–/– SMCs was significantly less than value from wild-type SMCs. ( b ) Number of SMCs per well after plating on 100 nM type I collagen and growing in DMEM containing 10% FCS for 1–4 days. Squares, DDR1+/+ SMCs; circles, DDR1–/– SMCs. Experiments were done in triplicate and repeated three times. A Value from DDR1–/– SMCs was significantly less than value from wild-type SMCs.

    Techniques Used: Mouse Assay, Incubation

    Related Articles

    Isolation:

    Article Title: Tbx1 regulates inherited metabolic and myogenic abilities of progenitor cells derived from slow- and fast-type muscle
    Article Snippet: The protein content was measured using a PierceTM BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA). .. Isolation and culture of single muscle fibersThe EDL, TA, and SOL muscles isolated from mice were digested with 2 mg/ml collagenase type I (Worthington Biochemical), as previously described [ ], and the isolated muscle fibers were suspended and cultured in growth medium (DMEM containing 15% horse serum [Invitrogen] with 0.5% chick embryo extract [Gemini Bio-Products, West Sacramento, CA] and penicillin–streptomycin) at 37 °C [ ]. .. EdU (Invitrogen) was added to the culture medium at a final concentration of 10 μM and cultured for 24 h.

    Article Title: VEGFR2 signaling drives meningeal vascular regeneration upon head injury
    Article Snippet: .. Magnetic-associated and fluorescence-activated cell sorting To sort dura mater or brain ECs, CD45+ hematopoietic or CD45− non-hematopoietic cells by FACS, tissues were isolated as described above and dissociated in collagenase type I (1 mg/ml, Worthington), dispase (1 mg/ml, Gibco), and DNase I (0.1 mg/ml, Merck) dissolved in DMEM/F12 supplemented with 5% fetal bovine serum (FBS) at 37 °C for 30 min. .. Cell suspensions went on RBC lysis by suspension in ACK lysis buffer (Gibco) for 5 min on ice and blocked with mouse anti-CD16/CD32 (553141, BD Bioscience) before being incubated for 20 min with indicated antibodies in FACS buffer (2% FBS in PBS).

    Mouse Assay:

    Article Title: Tbx1 regulates inherited metabolic and myogenic abilities of progenitor cells derived from slow- and fast-type muscle
    Article Snippet: The protein content was measured using a PierceTM BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA). .. Isolation and culture of single muscle fibersThe EDL, TA, and SOL muscles isolated from mice were digested with 2 mg/ml collagenase type I (Worthington Biochemical), as previously described [ ], and the isolated muscle fibers were suspended and cultured in growth medium (DMEM containing 15% horse serum [Invitrogen] with 0.5% chick embryo extract [Gemini Bio-Products, West Sacramento, CA] and penicillin–streptomycin) at 37 °C [ ]. .. EdU (Invitrogen) was added to the culture medium at a final concentration of 10 μM and cultured for 24 h.

    Cell Culture:

    Article Title: Tbx1 regulates inherited metabolic and myogenic abilities of progenitor cells derived from slow- and fast-type muscle
    Article Snippet: The protein content was measured using a PierceTM BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA). .. Isolation and culture of single muscle fibersThe EDL, TA, and SOL muscles isolated from mice were digested with 2 mg/ml collagenase type I (Worthington Biochemical), as previously described [ ], and the isolated muscle fibers were suspended and cultured in growth medium (DMEM containing 15% horse serum [Invitrogen] with 0.5% chick embryo extract [Gemini Bio-Products, West Sacramento, CA] and penicillin–streptomycin) at 37 °C [ ]. .. EdU (Invitrogen) was added to the culture medium at a final concentration of 10 μM and cultured for 24 h.

    Fluorescence:

    Article Title: VEGFR2 signaling drives meningeal vascular regeneration upon head injury
    Article Snippet: .. Magnetic-associated and fluorescence-activated cell sorting To sort dura mater or brain ECs, CD45+ hematopoietic or CD45− non-hematopoietic cells by FACS, tissues were isolated as described above and dissociated in collagenase type I (1 mg/ml, Worthington), dispase (1 mg/ml, Gibco), and DNase I (0.1 mg/ml, Merck) dissolved in DMEM/F12 supplemented with 5% fetal bovine serum (FBS) at 37 °C for 30 min. .. Cell suspensions went on RBC lysis by suspension in ACK lysis buffer (Gibco) for 5 min on ice and blocked with mouse anti-CD16/CD32 (553141, BD Bioscience) before being incubated for 20 min with indicated antibodies in FACS buffer (2% FBS in PBS).

    FACS:

    Article Title: VEGFR2 signaling drives meningeal vascular regeneration upon head injury
    Article Snippet: .. Magnetic-associated and fluorescence-activated cell sorting To sort dura mater or brain ECs, CD45+ hematopoietic or CD45− non-hematopoietic cells by FACS, tissues were isolated as described above and dissociated in collagenase type I (1 mg/ml, Worthington), dispase (1 mg/ml, Gibco), and DNase I (0.1 mg/ml, Merck) dissolved in DMEM/F12 supplemented with 5% fetal bovine serum (FBS) at 37 °C for 30 min. .. Cell suspensions went on RBC lysis by suspension in ACK lysis buffer (Gibco) for 5 min on ice and blocked with mouse anti-CD16/CD32 (553141, BD Bioscience) before being incubated for 20 min with indicated antibodies in FACS buffer (2% FBS in PBS).

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    Worthington Biochemical collagenase type 1
    AAV6.2FF transduces both airway and alveolar epithelial cells. a Whole-mount X-gal staining of the lungs, nasal cavity and trachea 3 weeks post-vector administration from either 1 × PBS or AAV6.2FF-emGFP-nlsCre treated Rosa26-Flox/LacZ mice. b Representative nuclear fast red counterstained paraffin sections (4 μm) from X-gal stained lungs. LacZ-positive cells were found in the distal airway epithelium and alveoli following delivery of AAV6.2FF-CMV-emGFP-nlsCre. Morphologic criteria demonstrate that both AT2 (white arrows) and club cells (yellow arrow) are X-gal positive (Distal lung scale bars represent 100 μm, 50 μm, and 20 μm from left to right; Trachea scale bars represent 200 μm, 50 μm, and 20 μm from left to right). c 3D histograms from flow cytometry analysis of dissociated whole lungs harvested from transgenic SP-B mice 8 days after IT administration of 10 11 vg per mouse of AAV-Luc control ( n = 2), or 10 11 vg per mouse of AAV6.2FF-GFP (AAV-GFP; n = 4). The first set of histograms represent the total CD45 negative, EpCAM++ (high expressing) cell population (first peak), and the CD45 negative, EpCAM++ cells that are GFP positive (second peak) in each mouse. The second set of histograms are the rescaled CD45 negative, EpCAM++, GFP positive cell populations as indicated by the black box in the first histogram. The column graph represents the mean percentage of total CD45 negative, EpCAM++ cells that are GFP positive with SD ( P value = two-tailed Student’s t test, * P = 0.0127). d This column graph represents the mean percentage of different lung cell populations with SD that are GFP positive. CD45 positive staining represents hematopoietic cells, CD45 negative, EpCAM dim staining represents alveolar <t>type</t> 1 (AT1) epithelial cells, CD45 negative, EpCAM++ staining represents AT2 or ciliated cells (red circles and red box), CD45 negative, CD31 positive staining represents endothelial cells, and CD45 negative, EpCAM negative, CD31 negative represents all other cell types found in dissociated lung tissue ( n = 4 biologically independent animals per group). The source data for c and d have been provided as a Source Data file.
    Collagenase Type 1, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/collagenase type 1/product/Worthington Biochemical
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    collagenase type 1 - by Bioz Stars, 2021-04
    99/100 stars
      Buy from Supplier

    Image Search Results


    AAV6.2FF transduces both airway and alveolar epithelial cells. a Whole-mount X-gal staining of the lungs, nasal cavity and trachea 3 weeks post-vector administration from either 1 × PBS or AAV6.2FF-emGFP-nlsCre treated Rosa26-Flox/LacZ mice. b Representative nuclear fast red counterstained paraffin sections (4 μm) from X-gal stained lungs. LacZ-positive cells were found in the distal airway epithelium and alveoli following delivery of AAV6.2FF-CMV-emGFP-nlsCre. Morphologic criteria demonstrate that both AT2 (white arrows) and club cells (yellow arrow) are X-gal positive (Distal lung scale bars represent 100 μm, 50 μm, and 20 μm from left to right; Trachea scale bars represent 200 μm, 50 μm, and 20 μm from left to right). c 3D histograms from flow cytometry analysis of dissociated whole lungs harvested from transgenic SP-B mice 8 days after IT administration of 10 11 vg per mouse of AAV-Luc control ( n = 2), or 10 11 vg per mouse of AAV6.2FF-GFP (AAV-GFP; n = 4). The first set of histograms represent the total CD45 negative, EpCAM++ (high expressing) cell population (first peak), and the CD45 negative, EpCAM++ cells that are GFP positive (second peak) in each mouse. The second set of histograms are the rescaled CD45 negative, EpCAM++, GFP positive cell populations as indicated by the black box in the first histogram. The column graph represents the mean percentage of total CD45 negative, EpCAM++ cells that are GFP positive with SD ( P value = two-tailed Student’s t test, * P = 0.0127). d This column graph represents the mean percentage of different lung cell populations with SD that are GFP positive. CD45 positive staining represents hematopoietic cells, CD45 negative, EpCAM dim staining represents alveolar type 1 (AT1) epithelial cells, CD45 negative, EpCAM++ staining represents AT2 or ciliated cells (red circles and red box), CD45 negative, CD31 positive staining represents endothelial cells, and CD45 negative, EpCAM negative, CD31 negative represents all other cell types found in dissociated lung tissue ( n = 4 biologically independent animals per group). The source data for c and d have been provided as a Source Data file.

    Journal: Nature Communications

    Article Title: A lung tropic AAV vector improves survival in a mouse model of surfactant B deficiency

    doi: 10.1038/s41467-020-17577-8

    Figure Lengend Snippet: AAV6.2FF transduces both airway and alveolar epithelial cells. a Whole-mount X-gal staining of the lungs, nasal cavity and trachea 3 weeks post-vector administration from either 1 × PBS or AAV6.2FF-emGFP-nlsCre treated Rosa26-Flox/LacZ mice. b Representative nuclear fast red counterstained paraffin sections (4 μm) from X-gal stained lungs. LacZ-positive cells were found in the distal airway epithelium and alveoli following delivery of AAV6.2FF-CMV-emGFP-nlsCre. Morphologic criteria demonstrate that both AT2 (white arrows) and club cells (yellow arrow) are X-gal positive (Distal lung scale bars represent 100 μm, 50 μm, and 20 μm from left to right; Trachea scale bars represent 200 μm, 50 μm, and 20 μm from left to right). c 3D histograms from flow cytometry analysis of dissociated whole lungs harvested from transgenic SP-B mice 8 days after IT administration of 10 11 vg per mouse of AAV-Luc control ( n = 2), or 10 11 vg per mouse of AAV6.2FF-GFP (AAV-GFP; n = 4). The first set of histograms represent the total CD45 negative, EpCAM++ (high expressing) cell population (first peak), and the CD45 negative, EpCAM++ cells that are GFP positive (second peak) in each mouse. The second set of histograms are the rescaled CD45 negative, EpCAM++, GFP positive cell populations as indicated by the black box in the first histogram. The column graph represents the mean percentage of total CD45 negative, EpCAM++ cells that are GFP positive with SD ( P value = two-tailed Student’s t test, * P = 0.0127). d This column graph represents the mean percentage of different lung cell populations with SD that are GFP positive. CD45 positive staining represents hematopoietic cells, CD45 negative, EpCAM dim staining represents alveolar type 1 (AT1) epithelial cells, CD45 negative, EpCAM++ staining represents AT2 or ciliated cells (red circles and red box), CD45 negative, CD31 positive staining represents endothelial cells, and CD45 negative, EpCAM negative, CD31 negative represents all other cell types found in dissociated lung tissue ( n = 4 biologically independent animals per group). The source data for c and d have been provided as a Source Data file.

    Article Snippet: Following perfusion, an enzyme mix containing 30 U neutral protease (Worthington; LS02104), 2500 U Collagenase I (Worthington; LS004196), 10 μg DNase I (Sigma; D5025-150KU) in DPBS with Mg2+ and Ca2+ was instilled into the lungs.

    Techniques: Staining, Plasmid Preparation, Mouse Assay, Flow Cytometry, Transgenic Assay, Expressing, Two Tailed Test

    [A] Digestion of BAT with collagenase I produces an adipose fraction harboring brown adipocytes and a stromal fraction composed primarily of endothelium separated by a fluid portion. [B] Deep-red fluorescence intensity images of centrifugation tubes containing

    Journal: Journal of materials chemistry. B, Materials for biology and medicine

    Article Title: Fluorescence Imaging of Interscapular Brown Adipose Tissue in Living Mice †

    doi: 10.1039/C4TB01914H

    Figure Lengend Snippet: [A] Digestion of BAT with collagenase I produces an adipose fraction harboring brown adipocytes and a stromal fraction composed primarily of endothelium separated by a fluid portion. [B] Deep-red fluorescence intensity images of centrifugation tubes containing

    Article Snippet: The minced tissue was digested with 2 mg/ml Collagenase I (Worthington Biochemical Corporation) in Hanks' Balanced Salt Solution (HBSS) (1.3 mM CaCl2 , 0.5 mM MgCl2 •6H2 O, 0.5 mM MgSO4 •7H2 O, 5.3 mM KCl, 0.40 mM KH2 PO4 , 4.2 mM NaHCO3 , 140 mM, NaCl, 0.3 mM Na2 HPO4 , pH 7.5) at 37 °C for 2 hours.

    Techniques: Fluorescence, Centrifugation

    Differences in the capacity of myogenic cells from slow- and fast-type muscles. a Freshly isolated intact myofibers from EDL or SOL muscle of wild-type mice were stained against Pax7 (red) and with DAPI (blue). Typical Pax7(+) and DAPI(+) cells are shown (arrowheads). The number of Pax7(+) satellite cells per fiber was quantified. Values are presented as mean ± SE ( n = 3). Scale bar = 50 µm. b Myofibers isolated from SOL or EDL muscle were cultured under growth conditions for 48 h. The activated satellite cells that migrated from each muscle fiber (arrowheads) were counted. Values are presented as mean ± SE ( n = 3). Scale bar = 100 µm. c Myoblasts from SOL or TA muscle were cultured in growth medium with EdU. After staining with EdU (green) and DAPI (blue), the proportion of EdU(+) cells among the total nuclei was counted. Values are presented as mean ± SE ( n = 3). Scale bar = 100 µm. d Myoblasts from SOL or TA muscle were cultured in differentiation medium for 4 days, and then myotubes and nuclei were stained against MyHC (green) and with DAPI (blue). The proportion of MyHC(+) cells among the total nuclei was quantified. Values are presented as mean ± SE ( n = 3). Scale bar = 50 µm. e Myotubes of TA- and SOL-derived myoblasts were stained against Myh7 (MyHC type I slow; green) and with DAPI (blue). Scale bar = 200 µm. f The expression levels of Myh7 , Myh2 , Myh1 , and Myh4 in myotubes were quantified by qPCR. The expression values were normalized to GAPDH expression and are presented as mean ± SE ( n = 4). g TA- or SOL-MBs derived from GFP mouse muscles were injected into TA muscles of NOD/ scid mice. Typical phase image (left) and GFP expression (right) are presented. Scale bar = 2 mm. h Longitudinal sections of engrafted TA muscles stained against slow-type myosin heavy chain (red) and laminin-α2 (gray), and with DAPI (blue). Scale bar = 500 µm. i OCR in SOL- or TA-derived myotubes was measured after treatment with oligomycin and FCCP. Values are presented as mean ± SE ( n = 3). j Basal OCR and SRC of myotubes were quantified. Values are presented as mean ± SE ( n = 3). k The expression levels of PGC-1 α, Nrf1 , and Tfam in TA- and SOL-MTs were quantified by qPCR. Values are presented as mean ± SE ( n = 3)

    Journal: Cell Death and Differentiation

    Article Title: Tbx1 regulates inherited metabolic and myogenic abilities of progenitor cells derived from slow- and fast-type muscle

    doi: 10.1038/s41418-018-0186-4

    Figure Lengend Snippet: Differences in the capacity of myogenic cells from slow- and fast-type muscles. a Freshly isolated intact myofibers from EDL or SOL muscle of wild-type mice were stained against Pax7 (red) and with DAPI (blue). Typical Pax7(+) and DAPI(+) cells are shown (arrowheads). The number of Pax7(+) satellite cells per fiber was quantified. Values are presented as mean ± SE ( n = 3). Scale bar = 50 µm. b Myofibers isolated from SOL or EDL muscle were cultured under growth conditions for 48 h. The activated satellite cells that migrated from each muscle fiber (arrowheads) were counted. Values are presented as mean ± SE ( n = 3). Scale bar = 100 µm. c Myoblasts from SOL or TA muscle were cultured in growth medium with EdU. After staining with EdU (green) and DAPI (blue), the proportion of EdU(+) cells among the total nuclei was counted. Values are presented as mean ± SE ( n = 3). Scale bar = 100 µm. d Myoblasts from SOL or TA muscle were cultured in differentiation medium for 4 days, and then myotubes and nuclei were stained against MyHC (green) and with DAPI (blue). The proportion of MyHC(+) cells among the total nuclei was quantified. Values are presented as mean ± SE ( n = 3). Scale bar = 50 µm. e Myotubes of TA- and SOL-derived myoblasts were stained against Myh7 (MyHC type I slow; green) and with DAPI (blue). Scale bar = 200 µm. f The expression levels of Myh7 , Myh2 , Myh1 , and Myh4 in myotubes were quantified by qPCR. The expression values were normalized to GAPDH expression and are presented as mean ± SE ( n = 4). g TA- or SOL-MBs derived from GFP mouse muscles were injected into TA muscles of NOD/ scid mice. Typical phase image (left) and GFP expression (right) are presented. Scale bar = 2 mm. h Longitudinal sections of engrafted TA muscles stained against slow-type myosin heavy chain (red) and laminin-α2 (gray), and with DAPI (blue). Scale bar = 500 µm. i OCR in SOL- or TA-derived myotubes was measured after treatment with oligomycin and FCCP. Values are presented as mean ± SE ( n = 3). j Basal OCR and SRC of myotubes were quantified. Values are presented as mean ± SE ( n = 3). k The expression levels of PGC-1 α, Nrf1 , and Tfam in TA- and SOL-MTs were quantified by qPCR. Values are presented as mean ± SE ( n = 3)

    Article Snippet: Isolation and culture of single muscle fibersThe EDL, TA, and SOL muscles isolated from mice were digested with 2 mg/ml collagenase type I (Worthington Biochemical), as previously described [ ], and the isolated muscle fibers were suspended and cultured in growth medium (DMEM containing 15% horse serum [Invitrogen] with 0.5% chick embryo extract [Gemini Bio-Products, West Sacramento, CA] and penicillin–streptomycin) at 37 °C [ ].

    Techniques: Isolation, Mouse Assay, Staining, Cell Culture, Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, Injection, Pyrolysis Gas Chromatography

    Dura mater ECs are superior to brain ECs in sprouting activity and show high expression of angiogenic molecules. a , b Images and comparisons of the number of sprouts (length, > 50 μm) of EC spheroids derived from FACS-sorted primary ECs of dura mater (DM) and brain (B) at 12 hr after incubation in collagen type I gel. Each dot indicates number of sprouts per spheroid. Each n = 6 from two independent experiments. Vertical bars indicate mean ± SD. P = 0.0022 versus brain ECs by two-tailed Mann–Whitney U test. Scale bars, 100 μm. c Highly magnified images of the left two panels in a showing robust sprouting (white arrowheads) and filopodia (yellow arrowheads) in EC spheroid from DM, whereas no sprouting and filopodia are seen in EC spheroid from B. Scale bars, 50 μm. d MA plot of significantly high-(1020 genes, red) and low (323 genes, blue)-expressing genes in dura mater ECs compared with brain ECs. Dotted horizontal lines demarcate twofold change between dura mater ECs and brain ECs. e IPA of 1343 differentially expressed genes in dura mater ECs compared with brain ECs. Ranked according to activation z score within Cardiovascular System Development and Function category. Vertical red-dotted line marks activation z score threshold. P

    Journal: Nature Communications

    Article Title: VEGFR2 signaling drives meningeal vascular regeneration upon head injury

    doi: 10.1038/s41467-020-17545-2

    Figure Lengend Snippet: Dura mater ECs are superior to brain ECs in sprouting activity and show high expression of angiogenic molecules. a , b Images and comparisons of the number of sprouts (length, > 50 μm) of EC spheroids derived from FACS-sorted primary ECs of dura mater (DM) and brain (B) at 12 hr after incubation in collagen type I gel. Each dot indicates number of sprouts per spheroid. Each n = 6 from two independent experiments. Vertical bars indicate mean ± SD. P = 0.0022 versus brain ECs by two-tailed Mann–Whitney U test. Scale bars, 100 μm. c Highly magnified images of the left two panels in a showing robust sprouting (white arrowheads) and filopodia (yellow arrowheads) in EC spheroid from DM, whereas no sprouting and filopodia are seen in EC spheroid from B. Scale bars, 50 μm. d MA plot of significantly high-(1020 genes, red) and low (323 genes, blue)-expressing genes in dura mater ECs compared with brain ECs. Dotted horizontal lines demarcate twofold change between dura mater ECs and brain ECs. e IPA of 1343 differentially expressed genes in dura mater ECs compared with brain ECs. Ranked according to activation z score within Cardiovascular System Development and Function category. Vertical red-dotted line marks activation z score threshold. P

    Article Snippet: Magnetic-associated and fluorescence-activated cell sorting To sort dura mater or brain ECs, CD45+ hematopoietic or CD45− non-hematopoietic cells by FACS, tissues were isolated as described above and dissociated in collagenase type I (1 mg/ml, Worthington), dispase (1 mg/ml, Gibco), and DNase I (0.1 mg/ml, Merck) dissolved in DMEM/F12 supplemented with 5% fetal bovine serum (FBS) at 37 °C for 30 min.

    Techniques: Activity Assay, Expressing, Derivative Assay, FACS, Incubation, Two Tailed Test, MANN-WHITNEY, Indirect Immunoperoxidase Assay, Activation Assay