recombinant human fibroblast growth factor basic Search Results


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  • 99
    Thermo Fisher fgf 2
    <t>FGF-2</t> induces HA accumulation in primary RPE cells. ( A – D ) FGF-2 induced HA accumulation in primary porcine RPE cells in a dose-dependent manner ( A ) 0 ng/mL, ( B ) 10 ng/mL, ( C ) 25 ng/mL, ( D ) 100 ng/mL. Fluorescence intensity was quantitated by integrated density measurement ( I ) (n ≥ 4, for each cell line). Data are presented as mean ± SEM ( E – H ) Z-plane images show that increased concentrations of FGF-2 induce increased apical accumulation of HA in addition to some peri-cellular and basal deposits of HA at high doses of FGF-2. Green: HA, red: phalloidin; blue: DAPI.
    Fgf 2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1005 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore fgf 2
    Neurite ingrowth with bridge at 6 weeks post implantation. A) Neurofilament stain (brown) of bridge implanted with the highest <t>VEGF/FGF-2</t> protein dose. The red dotted line marks the contours of the bridge at the implant site. Scale bar: 500 µm.
    Fgf 2, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 927 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher fgf basic aa 1 155 recombinant human protein
    Neurite ingrowth with bridge at 6 weeks post implantation. A) Neurofilament stain (brown) of bridge implanted with the highest <t>VEGF/FGF-2</t> protein dose. The red dotted line marks the contours of the bridge at the implant site. Scale bar: 500 µm.
    Fgf Basic Aa 1 155 Recombinant Human Protein, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 237 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher human basic fibroblast growth factor
    Neurite ingrowth with bridge at 6 weeks post implantation. A) Neurofilament stain (brown) of bridge implanted with the highest <t>VEGF/FGF-2</t> protein dose. The red dotted line marks the contours of the bridge at the implant site. Scale bar: 500 µm.
    Human Basic Fibroblast Growth Factor, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 399 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher human recombinant basic fibroblast growth factor
    Cancer upregulated drug-resistant (CUDR) enhances the embryonic stem cells (ESC) diffentiation into the hepatocyte-like cells . ( a ) Nuclear run-on ( upper ) and reverse-transcription polymerase chain reaction (RT-PCR) ( lower ) analysis of CUDR in <t>human</t> ESC line MEL-2 transfected with pCMV-A-GFP-CUDR and pCMA6-A-GFP respectively. β-actin as internal control. ( b ) Human ESC line MEL-2 transfected with pCMV-A-GFP-CUDR or pCMA6-A-GFP could efficiently generate definitive endoderm (DE) tissue by treating the the modified cultures with high concentrations of the TGFβ family ligand activin A (100 ng/ml) for 5 days. Western blot analysis with anti-Foxa2, anti-Sox17 in these induced cells (the 2nd, 3rd, and 5th day). β-actin as internal control. ( c ) A number of groups have generated hepatoblasts using this DE tissue as a starting material, plating the DE on matrix to mimic the hepatic ECM and then added FGF4 (100 ng/ml) and BMP (100 ng/ml) to mimic hepatic induction for 5 days. Western blotting with anti-AFP, anti-Albumin, and anti-HNF4α in the induced cells (the 6th and 10th day). β-actin as internal control. ( d ) Some combination of insulin-transferrinselenite (ITS, 5 μg/ml), hepatocyte <t>growth</t> <t>factor</t> (HGF) (20 ng/ml), oncostatin M (OSM) (10 ng/ml), acid <t>fibroblast</t> growth factor (aFGF) (50 ng/ml), and dexamethasone (10 −7 M) to expand the hepatoblast population and to promote hepatic maturation for 10 days. Western blotting with anti-Albumin in the induced cells (the 11th, 14th, 17th, and 20th day). β-actin as internal control. ( e ) Albumin promoter luciferase activity assay in derived hepatocyte-like cells. Each value was presented as mean ± standard error of the mean (SEM). ** P
    Human Recombinant Basic Fibroblast Growth Factor, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 357 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human recombinant basic fibroblast growth factor/product/Thermo Fisher
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    99
    PeproTech fgf 2
    <t>FGF‐2,</t> EGF, and PDGF‐BB reduce BMP‐2‐induced osteogenic differentiation of calvaria‐derived MPCs in vitro. In vitro expanded mouse calvarial MPCs were encapsulated in TG‐PEG hydrogels at a concentration of 1.5 × 10 6 mL −1 and cultured for 10 days in the presence of A,C) 0 and B,D) 100 ng mL −1 BMP‐2 as well as 50 ng mL −1 of the indicated growth factors ( n = 3). A,B) Colorimetric assessment of alkaline phosphatase activity (scale bars = 1 mm). C,D) Gene expression analysis of early markers of osteogenesis. Data are depicted as mean ± SD for all panels, * p
    Fgf 2, supplied by PeproTech, used in various techniques. Bioz Stars score: 99/100, based on 2110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Thermo Fisher fgf basic aa 10 155 recombinant human protein
    <t>FGF‐2,</t> EGF, and PDGF‐BB reduce BMP‐2‐induced osteogenic differentiation of calvaria‐derived MPCs in vitro. In vitro expanded mouse calvarial MPCs were encapsulated in TG‐PEG hydrogels at a concentration of 1.5 × 10 6 mL −1 and cultured for 10 days in the presence of A,C) 0 and B,D) 100 ng mL −1 BMP‐2 as well as 50 ng mL −1 of the indicated growth factors ( n = 3). A,B) Colorimetric assessment of alkaline phosphatase activity (scale bars = 1 mm). C,D) Gene expression analysis of early markers of osteogenesis. Data are depicted as mean ± SD for all panels, * p
    Fgf Basic Aa 10 155 Recombinant Human Protein, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 248 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    PeproTech recombinant human basic fibroblast growth factor
    <t>Basic</t> <t>fibroblast</t> <t>growth</t> <t>factor</t> ( bFGF) knockdown inhibits paracrine signaling‐mediated activation of endothelial cell proliferation and tube assembly in HDAC 1‐depleted CMC s. A, Immunoblots evaluating the expression of bFGF in total protein extracts derived from sh RNA ‐transduced CMC s (n=4). B, Densitometric analysis of bFGF immunoblots (expression relative to β‐actin). Values are mean± SEM (n=4). Western blot data were log base 10 (y=log 10 y) transformed and analyzed by unpaired, 1‐way ANOVA . P values were calculated using the post hoc Holm–Sidak multiple comparison test. C, HAEC growth was assessed following their propagation in CM from all sh RNA ‐transduced CMC groups (untransduced, sh NT , sh HDAC 1, shb FGF , and sh HDAC 1+shb FGF ). Values are mean± SEM (n=11). Growth data were analyzed by 1‐way ANOVA and P values determined using the post hoc Dunnett multiple comparison test. D, Representative fluorescent microscopy images (scale=1000 μm) and (E) graph enumerating HUVEC tube formation in response to incubation with CM from untransduced, sh NT , sh HDAC 1, shb FGF , or sh HDAC 1+shb FGF transduced CMC s. HUVEC base (−Ctrl), CMC base (−Ctrl), and HUVEC base+inducer (low serum growth supplement [ LSGS ]; +Ctrl) controls are included. Values are mean± SEM (n=8 for control and experimental groups). Tube formation data were analyzed using 1‐way ANOVA and P values determined using the post hoc Dunnett multiple comparison test. All experiments utilized 1:3 sh RNA viral titer dilutions. CM indicates conditioned medium; CMC , cardiac mesenchymal stromal cell; HAEC , <t>human</t> aortic endothelial cell; HDAC 1, histone deacetylase 1; HUVEC , human umbilical vein endothelial cell; shb FGF , short hairpin RNA basic fibroblast growth factor; sh HDAC 1, short hairpin RNA ‐histone deacetylase 1; sh NT , short hairpin RNA nontarget; UT , untransduced.
    Recombinant Human Basic Fibroblast Growth Factor, supplied by PeproTech, used in various techniques. Bioz Stars score: 99/100, based on 374 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    R&D Systems recombinant human fgf basic 146 aa protein
    ENC spheroid culture. a) Protocol (days 12–15) for ENC spheroid formation. NB+N2+B27; NB/N2/B27, Neurobasal medium with N2 and B27 supplement; <t>FGF2,</t> Recombinant Human <t>FGF</t> Basic; CHIR, CHIR 99021. b) Phase contrast and SOX10::GFP reporter line GFP expression of 3D spheroids on day 14. Scale bar = 200 μm.
    Recombinant Human Fgf Basic 146 Aa Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 163 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    FUJIFILM recombinant human basic fibroblast growth factor
    ENC spheroid culture. a) Protocol (days 12–15) for ENC spheroid formation. NB+N2+B27; NB/N2/B27, Neurobasal medium with N2 and B27 supplement; <t>FGF2,</t> Recombinant Human <t>FGF</t> Basic; CHIR, CHIR 99021. b) Phase contrast and SOX10::GFP reporter line GFP expression of 3D spheroids on day 14. Scale bar = 200 μm.
    Recombinant Human Basic Fibroblast Growth Factor, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 93/100, based on 256 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant human basic fibroblast growth factor/product/FUJIFILM
    Average 93 stars, based on 256 article reviews
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    recombinant human basic fibroblast growth factor - by Bioz Stars, 2020-09
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    93
    Millipore recombinant human basic fibroblast growth factor
    ENC spheroid culture. a) Protocol (days 12–15) for ENC spheroid formation. NB+N2+B27; NB/N2/B27, Neurobasal medium with N2 and B27 supplement; <t>FGF2,</t> Recombinant Human <t>FGF</t> Basic; CHIR, CHIR 99021. b) Phase contrast and SOX10::GFP reporter line GFP expression of 3D spheroids on day 14. Scale bar = 200 μm.
    Recombinant Human Basic Fibroblast Growth Factor, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 136 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant human basic fibroblast growth factor/product/Millipore
    Average 93 stars, based on 136 article reviews
    Price from $9.99 to $1999.99
    recombinant human basic fibroblast growth factor - by Bioz Stars, 2020-09
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    99
    Millipore human recombinant basic fibroblast growth factor
    ENC spheroid culture. a) Protocol (days 12–15) for ENC spheroid formation. NB+N2+B27; NB/N2/B27, Neurobasal medium with N2 and B27 supplement; <t>FGF2,</t> Recombinant Human <t>FGF</t> Basic; CHIR, CHIR 99021. b) Phase contrast and SOX10::GFP reporter line GFP expression of 3D spheroids on day 14. Scale bar = 200 μm.
    Human Recombinant Basic Fibroblast Growth Factor, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 121 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human recombinant basic fibroblast growth factor/product/Millipore
    Average 99 stars, based on 121 article reviews
    Price from $9.99 to $1999.99
    human recombinant basic fibroblast growth factor - by Bioz Stars, 2020-09
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    94
    R&D Systems recombinant human fgf 2
    Intronic precursor of miR-4673 in notch-1 codes an active enhancer. a, Structural analysis shows improved thermodynamic stability of the RNA hairpin in primate lineage culminating in the structural maturation of miR-4673 hairpin in Hominins . b, Nucleosome-favouring dinucleotide usage map of human notch-1 intron 4. c, Transmission electron micrograph of a nucleosome formed by NPS miR (left). Gel retardation (middle) and restriction enzyme digestion assays (right) provided further confirmation for nucleosome formation by NPS miR . d , The temporal profile of enhancer-RNAs (eRNAs) that originates from Notch-1 intron-4 (full blots are provided in supplementary file 10). Vertical lines show the location of eRNAs with reference to the dinucleotide usage map. In order to access the eRNA profile, cells were synchronised at G0 and released into G1 by application of a single pulse of <t>FGF-2.</t> Note the temporal stability of 3′-eRNA from a region that corresponds to NPS miR (see supplementary file 11). This region is positioned at 3′-terminus of a stable RNA palindrome formed by two Alu elements. e, Histogram shows cumulative distribution of TCF3/4 and TFAP2A/B/C cis -motifs in the intron 4 of human notch-1. At the bottom, Levenshtein distance (L.E.D) as a measure of intronic change is aligned to the enhancer map. Sequences that belong to AluJb and AluYa1 insertions in all Simians are in grey. Species-specific transpositional events are marked in LED heat maps.
    Recombinant Human Fgf 2, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 99 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 99 article reviews
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    recombinant human fgf 2 - by Bioz Stars, 2020-09
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    Image Search Results


    FGF-2 induces HA accumulation in primary RPE cells. ( A – D ) FGF-2 induced HA accumulation in primary porcine RPE cells in a dose-dependent manner ( A ) 0 ng/mL, ( B ) 10 ng/mL, ( C ) 25 ng/mL, ( D ) 100 ng/mL. Fluorescence intensity was quantitated by integrated density measurement ( I ) (n ≥ 4, for each cell line). Data are presented as mean ± SEM ( E – H ) Z-plane images show that increased concentrations of FGF-2 induce increased apical accumulation of HA in addition to some peri-cellular and basal deposits of HA at high doses of FGF-2. Green: HA, red: phalloidin; blue: DAPI.

    Journal: Cells

    Article Title: Role of FGF and Hyaluronan in Choroidal Neovascularization in Sorsby Fundus Dystrophy

    doi: 10.3390/cells9030608

    Figure Lengend Snippet: FGF-2 induces HA accumulation in primary RPE cells. ( A – D ) FGF-2 induced HA accumulation in primary porcine RPE cells in a dose-dependent manner ( A ) 0 ng/mL, ( B ) 10 ng/mL, ( C ) 25 ng/mL, ( D ) 100 ng/mL. Fluorescence intensity was quantitated by integrated density measurement ( I ) (n ≥ 4, for each cell line). Data are presented as mean ± SEM ( E – H ) Z-plane images show that increased concentrations of FGF-2 induce increased apical accumulation of HA in addition to some peri-cellular and basal deposits of HA at high doses of FGF-2. Green: HA, red: phalloidin; blue: DAPI.

    Article Snippet: Cells were serum-starved for 24 h before treatment with the FGF Receptor inhibitor BGJ-398 (Selleckchem, Houston, TX, USA, #S2183) for 48 h. Similarly, cells were treated with FGF-2 (Gibco from Thermo Fisher Scientific, #13256-029) with the required cofactor heparin sodium salt (1 μg/mL, Sigma Aldrich, #H3149) for 48 h after serum starving for 24 h.

    Techniques: Fluorescence

    Growth factor stimulation of myocilin secretion from NTM cells Western blot of conditioned media collected from NTM cells treated for 72 hours with 1000 ng/ml (super physiologic concentrations) of recombinant human basic fibroblast growth factor (bFGF), transforming growth factor beta-2 (TGF-β2), epidermal growth factor (EGF), platelet-derived endothelial growth factor (PD-ECGF), endothelin-1(ET-1), vascular endothelial growth factor (VEGF) or osteopontin (OPN). UnCM = unconditioned media, CM = conditioned media.

    Journal: Experimental eye research

    Article Title: Aqueous Humor Rapidly Stimulates Myocilin Secretion from Human Trabecular Meshwork Cells

    doi: 10.1016/j.exer.2010.09.017

    Figure Lengend Snippet: Growth factor stimulation of myocilin secretion from NTM cells Western blot of conditioned media collected from NTM cells treated for 72 hours with 1000 ng/ml (super physiologic concentrations) of recombinant human basic fibroblast growth factor (bFGF), transforming growth factor beta-2 (TGF-β2), epidermal growth factor (EGF), platelet-derived endothelial growth factor (PD-ECGF), endothelin-1(ET-1), vascular endothelial growth factor (VEGF) or osteopontin (OPN). UnCM = unconditioned media, CM = conditioned media.

    Article Snippet: Cells were treated for 72 hours with 10, 100 or 1000 ng/ml of the following growth factors: recombinant human basic fibroblast growth factor (bFGF; GIBCO), transforming growth factor beta-2 (TGF-β2) (R & D Systems, Minneapolis, MN), epidermal growth factor (EGF) (GIBCO), platelet-derived endothelial growth factor (PD-ECGF) (R & D Systems), endothelin-1 (ET-1) (Sigma-Aldrich), or osteopontin (R & D Systems).

    Techniques: Western Blot, Recombinant, Derivative Assay

    Effect of LIF and bFGF on endogenous genes: REX-1 (a), NANOG (b), OCT4 (c), and SOX2 (d) genes. Results expressed as the mean of fold change ±2 standard deviation (SD) relative to control (LIF and bFGF). ∗ p

    Journal: Stem Cells International

    Article Title: Inhibition of JAK-STAT ERK/MAPK and Glycogen Synthase Kinase-3 Induces a Change in Gene Expression Profile of Bovine Induced Pluripotent Stem Cells

    doi: 10.1155/2016/5127984

    Figure Lengend Snippet: Effect of LIF and bFGF on endogenous genes: REX-1 (a), NANOG (b), OCT4 (c), and SOX2 (d) genes. Results expressed as the mean of fold change ±2 standard deviation (SD) relative to control (LIF and bFGF). ∗ p

    Article Snippet: The biPSC media consisted of Minimum Essential Medium Alpha (MEM-α ) with L-glutamine ribonucleosides and deoxyribonucleosides, Fetal Calf Serum 20% (JRH Bioproducts), GlutaMAX 2 mM, Nonessential Amino Acids (NEAA) 10 μ M (Gibco), Human recombinant LIF 5 ng/mL (Sigma), recombinant human Basic Fibroblast Growth Factor (bFGF) 10 ng/mL (Invitrogen), 2-Mercaptoethanol 55 μ M, and Penicillin-Streptomycin (25 units and 25 μ g, resp., Invitrogen).

    Techniques: Standard Deviation

    Neurite ingrowth with bridge at 6 weeks post implantation. A) Neurofilament stain (brown) of bridge implanted with the highest VEGF/FGF-2 protein dose. The red dotted line marks the contours of the bridge at the implant site. Scale bar: 500 µm.

    Journal: Journal of biomedical materials research. Part A

    Article Title: VEGF and FGF-2 delivery from spinal cord bridges to enhance angiogenesis following injury

    doi: 10.1002/jbm.a.33112

    Figure Lengend Snippet: Neurite ingrowth with bridge at 6 weeks post implantation. A) Neurofilament stain (brown) of bridge implanted with the highest VEGF/FGF-2 protein dose. The red dotted line marks the contours of the bridge at the implant site. Scale bar: 500 µm.

    Article Snippet: Proteins were encapsulated into the microspheres by emulsifying a protein solution (17 µl), containing VEGF (recombinant mouse VEGF-165, Prospec, Rehovot, Israel) or FGF-2 (recombinant human FGF-2, Chemicon, Billerica, Massachusetts), 700 µg bovine serum albumin (BSA), 50 mg/mL sucrose, and MgCO3 (3% wt of BSA), with 500 µL 3 wt% PLG in dichloromethane using sonication at 40 watt for 15 s on ice.

    Techniques: Staining

    Protein release from bridge. Bridges were fabricated with VEGF (filled line) or FGF-2 (dotted line) in three different manners: mixing only (squares), encapsulation only (triangles), or a combination of both encapsulation and mixing (circles). The amount

    Journal: Journal of biomedical materials research. Part A

    Article Title: VEGF and FGF-2 delivery from spinal cord bridges to enhance angiogenesis following injury

    doi: 10.1002/jbm.a.33112

    Figure Lengend Snippet: Protein release from bridge. Bridges were fabricated with VEGF (filled line) or FGF-2 (dotted line) in three different manners: mixing only (squares), encapsulation only (triangles), or a combination of both encapsulation and mixing (circles). The amount

    Article Snippet: Proteins were encapsulated into the microspheres by emulsifying a protein solution (17 µl), containing VEGF (recombinant mouse VEGF-165, Prospec, Rehovot, Israel) or FGF-2 (recombinant human FGF-2, Chemicon, Billerica, Massachusetts), 700 µg bovine serum albumin (BSA), 50 mg/mL sucrose, and MgCO3 (3% wt of BSA), with 500 µL 3 wt% PLG in dichloromethane using sonication at 40 watt for 15 s on ice.

    Techniques:

    VEGF and FGF-2 activity during in vitro release. Phosphorylation of (A) VEGF and (B) FGF-2 receptors was assessed by western blot. Legend: 1. control (100 ng/ml VEGF or 50 ng/ml FGF-2), 2. mixing 24 hours, 3. encapsulation 24 hours, 4. mixing 5 days,

    Journal: Journal of biomedical materials research. Part A

    Article Title: VEGF and FGF-2 delivery from spinal cord bridges to enhance angiogenesis following injury

    doi: 10.1002/jbm.a.33112

    Figure Lengend Snippet: VEGF and FGF-2 activity during in vitro release. Phosphorylation of (A) VEGF and (B) FGF-2 receptors was assessed by western blot. Legend: 1. control (100 ng/ml VEGF or 50 ng/ml FGF-2), 2. mixing 24 hours, 3. encapsulation 24 hours, 4. mixing 5 days,

    Article Snippet: Proteins were encapsulated into the microspheres by emulsifying a protein solution (17 µl), containing VEGF (recombinant mouse VEGF-165, Prospec, Rehovot, Israel) or FGF-2 (recombinant human FGF-2, Chemicon, Billerica, Massachusetts), 700 µg bovine serum albumin (BSA), 50 mg/mL sucrose, and MgCO3 (3% wt of BSA), with 500 µL 3 wt% PLG in dichloromethane using sonication at 40 watt for 15 s on ice.

    Techniques: Activity Assay, In Vitro, Western Blot

    Endothelial cell infiltration in bridge at 6 weeks post implantation. A) RECA stain (brown): from left to right: bridge containing 4 µg of VEGF encapsulated within microspheres and 2 µg of FGF-2 and VEGF mixed (high protein), and bridge

    Journal: Journal of biomedical materials research. Part A

    Article Title: VEGF and FGF-2 delivery from spinal cord bridges to enhance angiogenesis following injury

    doi: 10.1002/jbm.a.33112

    Figure Lengend Snippet: Endothelial cell infiltration in bridge at 6 weeks post implantation. A) RECA stain (brown): from left to right: bridge containing 4 µg of VEGF encapsulated within microspheres and 2 µg of FGF-2 and VEGF mixed (high protein), and bridge

    Article Snippet: Proteins were encapsulated into the microspheres by emulsifying a protein solution (17 µl), containing VEGF (recombinant mouse VEGF-165, Prospec, Rehovot, Israel) or FGF-2 (recombinant human FGF-2, Chemicon, Billerica, Massachusetts), 700 µg bovine serum albumin (BSA), 50 mg/mL sucrose, and MgCO3 (3% wt of BSA), with 500 µL 3 wt% PLG in dichloromethane using sonication at 40 watt for 15 s on ice.

    Techniques: Staining

    Protein encapsulation efficiency and protein retention after leaching. A) VEGF and FGF-2 encapsulation efficiency after microsphere encapsulation and collection. B) VEGF and FGF-2 retention after leaching bridges for 1 h. Proteins were incorporated using

    Journal: Journal of biomedical materials research. Part A

    Article Title: VEGF and FGF-2 delivery from spinal cord bridges to enhance angiogenesis following injury

    doi: 10.1002/jbm.a.33112

    Figure Lengend Snippet: Protein encapsulation efficiency and protein retention after leaching. A) VEGF and FGF-2 encapsulation efficiency after microsphere encapsulation and collection. B) VEGF and FGF-2 retention after leaching bridges for 1 h. Proteins were incorporated using

    Article Snippet: Proteins were encapsulated into the microspheres by emulsifying a protein solution (17 µl), containing VEGF (recombinant mouse VEGF-165, Prospec, Rehovot, Israel) or FGF-2 (recombinant human FGF-2, Chemicon, Billerica, Massachusetts), 700 µg bovine serum albumin (BSA), 50 mg/mL sucrose, and MgCO3 (3% wt of BSA), with 500 µL 3 wt% PLG in dichloromethane using sonication at 40 watt for 15 s on ice.

    Techniques:

    Cancer upregulated drug-resistant (CUDR) enhances the embryonic stem cells (ESC) diffentiation into the hepatocyte-like cells . ( a ) Nuclear run-on ( upper ) and reverse-transcription polymerase chain reaction (RT-PCR) ( lower ) analysis of CUDR in human ESC line MEL-2 transfected with pCMV-A-GFP-CUDR and pCMA6-A-GFP respectively. β-actin as internal control. ( b ) Human ESC line MEL-2 transfected with pCMV-A-GFP-CUDR or pCMA6-A-GFP could efficiently generate definitive endoderm (DE) tissue by treating the the modified cultures with high concentrations of the TGFβ family ligand activin A (100 ng/ml) for 5 days. Western blot analysis with anti-Foxa2, anti-Sox17 in these induced cells (the 2nd, 3rd, and 5th day). β-actin as internal control. ( c ) A number of groups have generated hepatoblasts using this DE tissue as a starting material, plating the DE on matrix to mimic the hepatic ECM and then added FGF4 (100 ng/ml) and BMP (100 ng/ml) to mimic hepatic induction for 5 days. Western blotting with anti-AFP, anti-Albumin, and anti-HNF4α in the induced cells (the 6th and 10th day). β-actin as internal control. ( d ) Some combination of insulin-transferrinselenite (ITS, 5 μg/ml), hepatocyte growth factor (HGF) (20 ng/ml), oncostatin M (OSM) (10 ng/ml), acid fibroblast growth factor (aFGF) (50 ng/ml), and dexamethasone (10 −7 M) to expand the hepatoblast population and to promote hepatic maturation for 10 days. Western blotting with anti-Albumin in the induced cells (the 11th, 14th, 17th, and 20th day). β-actin as internal control. ( e ) Albumin promoter luciferase activity assay in derived hepatocyte-like cells. Each value was presented as mean ± standard error of the mean (SEM). ** P

    Journal: Molecular Therapy

    Article Title: Long Noncoding RNA CUDR Regulates HULC and β-Catenin to Govern Human Liver Stem Cell Malignant Differentiation

    doi: 10.1038/mt.2015.166

    Figure Lengend Snippet: Cancer upregulated drug-resistant (CUDR) enhances the embryonic stem cells (ESC) diffentiation into the hepatocyte-like cells . ( a ) Nuclear run-on ( upper ) and reverse-transcription polymerase chain reaction (RT-PCR) ( lower ) analysis of CUDR in human ESC line MEL-2 transfected with pCMV-A-GFP-CUDR and pCMA6-A-GFP respectively. β-actin as internal control. ( b ) Human ESC line MEL-2 transfected with pCMV-A-GFP-CUDR or pCMA6-A-GFP could efficiently generate definitive endoderm (DE) tissue by treating the the modified cultures with high concentrations of the TGFβ family ligand activin A (100 ng/ml) for 5 days. Western blot analysis with anti-Foxa2, anti-Sox17 in these induced cells (the 2nd, 3rd, and 5th day). β-actin as internal control. ( c ) A number of groups have generated hepatoblasts using this DE tissue as a starting material, plating the DE on matrix to mimic the hepatic ECM and then added FGF4 (100 ng/ml) and BMP (100 ng/ml) to mimic hepatic induction for 5 days. Western blotting with anti-AFP, anti-Albumin, and anti-HNF4α in the induced cells (the 6th and 10th day). β-actin as internal control. ( d ) Some combination of insulin-transferrinselenite (ITS, 5 μg/ml), hepatocyte growth factor (HGF) (20 ng/ml), oncostatin M (OSM) (10 ng/ml), acid fibroblast growth factor (aFGF) (50 ng/ml), and dexamethasone (10 −7 M) to expand the hepatoblast population and to promote hepatic maturation for 10 days. Western blotting with anti-Albumin in the induced cells (the 11th, 14th, 17th, and 20th day). β-actin as internal control. ( e ) Albumin promoter luciferase activity assay in derived hepatocyte-like cells. Each value was presented as mean ± standard error of the mean (SEM). ** P

    Article Snippet: Cells were collected and washed to remove serum; then suspended in serum-free Dulbecco's Modified Eagle Medium/F12 supplemented with 20 ng/ml human recombinant epidermal growth factor, 10 ng/ml human recombinant basic fibroblast growth factor, 2% B27 supplement without vitamin A, and 1% N2 supplement (Invitrogen).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Transfection, Modification, Western Blot, Generated, Luciferase, Activity Assay, Derivative Assay

    Culturing of bone marrow cells isolated from adult JCV T-antigen transgenic mice. Mesenchymal stem cells (MSCs) were isolated from the bone marrow of adult JCV T-antigen transgenic mice by the virtue of plastic adherence when cultured in mesenchymal media composed of α-MEM containing 20% fetal bovine serum. Non-adherent cells were removed and discarded while adherent cells were then transferred to neural stem cell media (containing bFGF and EGF) for 2–3 weeks or maintained in mesenchymal media. Cells in both culture conditions were monitored for growth and analyzed for expression of JCV T-antigen. Neural crest cells were further expanded and analyzed for expression of neural crest markers and differentiation into glial and osteogenic components to confirm their neural crest origin.

    Journal: PLoS ONE

    Article Title: Neural Crest Cells Isolated from the Bone Marrow of Transgenic Mice Express JCV T-Antigen

    doi: 10.1371/journal.pone.0065947

    Figure Lengend Snippet: Culturing of bone marrow cells isolated from adult JCV T-antigen transgenic mice. Mesenchymal stem cells (MSCs) were isolated from the bone marrow of adult JCV T-antigen transgenic mice by the virtue of plastic adherence when cultured in mesenchymal media composed of α-MEM containing 20% fetal bovine serum. Non-adherent cells were removed and discarded while adherent cells were then transferred to neural stem cell media (containing bFGF and EGF) for 2–3 weeks or maintained in mesenchymal media. Cells in both culture conditions were monitored for growth and analyzed for expression of JCV T-antigen. Neural crest cells were further expanded and analyzed for expression of neural crest markers and differentiation into glial and osteogenic components to confirm their neural crest origin.

    Article Snippet: After an additional four days in culture, cells were harvested with trypsin (0.25%) and cultured in standard mesenchymal media composed of α-MEM media supplemented with 20% FBS or incubated with serum-free Neurobasal media supplemented with 20 ηg/µL of epidermal growth factor (EGF) (Invitrogen) and 20 ηg/µL of basic fibroblast growth factor (bFGF) (Invitrogen) and B27 (1∶50; Invitrogen) for 2–3 weeks.

    Techniques: Isolation, Transgenic Assay, Mouse Assay, Cell Culture, Expressing

    FGF‐2, EGF, and PDGF‐BB reduce BMP‐2‐induced osteogenic differentiation of calvaria‐derived MPCs in vitro. In vitro expanded mouse calvarial MPCs were encapsulated in TG‐PEG hydrogels at a concentration of 1.5 × 10 6 mL −1 and cultured for 10 days in the presence of A,C) 0 and B,D) 100 ng mL −1 BMP‐2 as well as 50 ng mL −1 of the indicated growth factors ( n = 3). A,B) Colorimetric assessment of alkaline phosphatase activity (scale bars = 1 mm). C,D) Gene expression analysis of early markers of osteogenesis. Data are depicted as mean ± SD for all panels, * p

    Journal: Advanced Science

    Article Title: Smart Hydrogels for the Augmentation of Bone Regeneration by Endogenous Mesenchymal Progenitor Cell Recruitment, Smart Hydrogels for the Augmentation of Bone Regeneration by Endogenous Mesenchymal Progenitor Cell Recruitment

    doi: 10.1002/advs.201903395

    Figure Lengend Snippet: FGF‐2, EGF, and PDGF‐BB reduce BMP‐2‐induced osteogenic differentiation of calvaria‐derived MPCs in vitro. In vitro expanded mouse calvarial MPCs were encapsulated in TG‐PEG hydrogels at a concentration of 1.5 × 10 6 mL −1 and cultured for 10 days in the presence of A,C) 0 and B,D) 100 ng mL −1 BMP‐2 as well as 50 ng mL −1 of the indicated growth factors ( n = 3). A,B) Colorimetric assessment of alkaline phosphatase activity (scale bars = 1 mm). C,D) Gene expression analysis of early markers of osteogenesis. Data are depicted as mean ± SD for all panels, * p

    Article Snippet: Cell Culture Freshly sorted cells were cultured as adherent cultures on collagen‐coated plates at 37 °C with 5% CO2 in maintenance medium: Minimal essential medium alpha (MEMα; Gibco Life Technologies, cat. no. 22571‐020) with 10% v/v fetal calf serum (FCS; Gibco Life Technologies, cat. no. 10500) and 1% P/S supplemented with 5 ng mL−1 FGF‐2 (Peprotech, cat. no. 100‐18B).

    Techniques: Derivative Assay, In Vitro, Concentration Assay, Cell Culture, Activity Assay, Expressing

    EGF, FGF‐2, and PDGF‐BB induce 3D migration of Sca‐1 + MPCs in an in vitro growth factor and cytokine screen. Spheroids consisting of 750 prospectively isolated Sca‐1 + MPCs were embedded in TG‐PEG hydrogels and cultured for 72 h in the presence of murine biomolecules: 10 ng mL −1 EGF ( n = 5) and FGF‐2 ( n = 4); 20 ng mL −1 IL‐6 ( n = 5) and TNFα ( n = 9); 30 ng mL −1 IGF −1 ( n = 6); 50 ng mL −1 BMP‐2 ( n = 5), BMP‐4 ( n = 3), HGF ( n = 4), and SCF ( n = 5); 100 ng mL −1 PDGF‐BB ( n = 5) and VEGF 165 ( n = 5); 150 ng mL −1 CCL2 ( n = 7), CCL5 ( n = 5), CCL7 ( n = 6), CCL21 ( n = 4), CCL22 ( n = 6), CXCL10 ( n = 11), CXCL11 ( n = 5), CXCL12 ( n = 10), and CXCL13 ( n = 4). Collective cell migration (gray bars) and single‐cell migration (black) were quantified by automated image analysis. Data are depicted as mean ± SD. px, pixels. Scale bar = 200 µm.

    Journal: Advanced Science

    Article Title: Smart Hydrogels for the Augmentation of Bone Regeneration by Endogenous Mesenchymal Progenitor Cell Recruitment, Smart Hydrogels for the Augmentation of Bone Regeneration by Endogenous Mesenchymal Progenitor Cell Recruitment

    doi: 10.1002/advs.201903395

    Figure Lengend Snippet: EGF, FGF‐2, and PDGF‐BB induce 3D migration of Sca‐1 + MPCs in an in vitro growth factor and cytokine screen. Spheroids consisting of 750 prospectively isolated Sca‐1 + MPCs were embedded in TG‐PEG hydrogels and cultured for 72 h in the presence of murine biomolecules: 10 ng mL −1 EGF ( n = 5) and FGF‐2 ( n = 4); 20 ng mL −1 IL‐6 ( n = 5) and TNFα ( n = 9); 30 ng mL −1 IGF −1 ( n = 6); 50 ng mL −1 BMP‐2 ( n = 5), BMP‐4 ( n = 3), HGF ( n = 4), and SCF ( n = 5); 100 ng mL −1 PDGF‐BB ( n = 5) and VEGF 165 ( n = 5); 150 ng mL −1 CCL2 ( n = 7), CCL5 ( n = 5), CCL7 ( n = 6), CCL21 ( n = 4), CCL22 ( n = 6), CXCL10 ( n = 11), CXCL11 ( n = 5), CXCL12 ( n = 10), and CXCL13 ( n = 4). Collective cell migration (gray bars) and single‐cell migration (black) were quantified by automated image analysis. Data are depicted as mean ± SD. px, pixels. Scale bar = 200 µm.

    Article Snippet: Cell Culture Freshly sorted cells were cultured as adherent cultures on collagen‐coated plates at 37 °C with 5% CO2 in maintenance medium: Minimal essential medium alpha (MEMα; Gibco Life Technologies, cat. no. 22571‐020) with 10% v/v fetal calf serum (FCS; Gibco Life Technologies, cat. no. 10500) and 1% P/S supplemented with 5 ng mL−1 FGF‐2 (Peprotech, cat. no. 100‐18B).

    Techniques: Migration, In Vitro, Isolation, Cell Culture

    Wnt3A and Rspo2 maintains stemness in GSCs. a Rspo2 and Wnt3A upregulates stem cell marker in U251 GSCs as demonstrated by real time PCR. Blk indicates U251 GSCs cultured in GSC media with 0.1% DMSO. b All-trans retinoic acid (10 µM RA) was used to induce differentiation in U251 GSCs for 24 h with or without Wnt ligands (20 ng/ml). Real-time PCR was used to determine the effect on differentiation. Results show that Rspo2/Wnt3A treatment rescues RA-induced GSC differentiation. Blk indicates U251 GSCs cultured in DMEM with 0.1% DMSO. c GSCs were cultured in GSC media, or GSC media without EGF and FGF, or GSC media without EGF and FGF but with Wnt3A and Rspo2 for 7 days. Results show that lack of EGF/FGF causes reduced sphere formation, which can be compensated by adding Rspo2/Wnt3A. d Real-time PCR shows that Rspo2/Wnt3A treatment abolishes the downregulation of stem cell markers and upregulation of differentiation markers caused by growth factor deprivation. Blk in C and D indicates U251 GSCs cultured in GSC media with 0.1% DMSO

    Journal: Cancer Cell International

    Article Title: R-spodin2 enhances canonical Wnt signaling to maintain the stemness of glioblastoma cells

    doi: 10.1186/s12935-018-0655-3

    Figure Lengend Snippet: Wnt3A and Rspo2 maintains stemness in GSCs. a Rspo2 and Wnt3A upregulates stem cell marker in U251 GSCs as demonstrated by real time PCR. Blk indicates U251 GSCs cultured in GSC media with 0.1% DMSO. b All-trans retinoic acid (10 µM RA) was used to induce differentiation in U251 GSCs for 24 h with or without Wnt ligands (20 ng/ml). Real-time PCR was used to determine the effect on differentiation. Results show that Rspo2/Wnt3A treatment rescues RA-induced GSC differentiation. Blk indicates U251 GSCs cultured in DMEM with 0.1% DMSO. c GSCs were cultured in GSC media, or GSC media without EGF and FGF, or GSC media without EGF and FGF but with Wnt3A and Rspo2 for 7 days. Results show that lack of EGF/FGF causes reduced sphere formation, which can be compensated by adding Rspo2/Wnt3A. d Real-time PCR shows that Rspo2/Wnt3A treatment abolishes the downregulation of stem cell markers and upregulation of differentiation markers caused by growth factor deprivation. Blk in C and D indicates U251 GSCs cultured in GSC media with 0.1% DMSO

    Article Snippet: The following GSC culture medium was applied to enrich GSCs: DMEM/F12 medium (Thermo Fisher Scientific, Grand Island, NY) supplemented with 1 × B-27™ Supplement, serum free (Gibco, 17504044), 20 ng/ml Animal-Free Recombinant Human EGF (PeproTech, AF-100-15), 20 ng/ml Recombinant Human FGF-basic (154 a.a.) (PeproTech, 100-18B), and 1× penicillin–streptomycin.

    Techniques: Marker, Real-time Polymerase Chain Reaction, Cell Culture

    Model for the role of Nox2 in FGF-2 redox signaling in AHPs The mitogen FGF-2 induces the production of H 2 O 2 in AHPs, which can be blocked by either the general flavin inhibitor DPI, the antioxidant NAC, the expression of Catalase or genetic manipulation of Nox2. Nox2-generated H 2 O 2 oxidizes and deactivates PTEN, which enhances signaling through Akt and manifest phenotypes in growth rates of AHPs in vitro and in vivo .

    Journal: Nature chemical biology

    Article Title: Nox2 redox signaling maintains essential cell populations in the brain

    doi: 10.1038/nchembio.497

    Figure Lengend Snippet: Model for the role of Nox2 in FGF-2 redox signaling in AHPs The mitogen FGF-2 induces the production of H 2 O 2 in AHPs, which can be blocked by either the general flavin inhibitor DPI, the antioxidant NAC, the expression of Catalase or genetic manipulation of Nox2. Nox2-generated H 2 O 2 oxidizes and deactivates PTEN, which enhances signaling through Akt and manifest phenotypes in growth rates of AHPs in vitro and in vivo .

    Article Snippet: Cell Culture AHPs were cultured on tissue culture polystyrene coated with poly-ornithine and 5 μg/mL of laminin (Invitrogen) and grown in Dulbecco's modified Eagle medium (DMEM)/F-12 (1:1) high-glucose medium (Invitrogen) containing N-2 supplement (Invitrogen) and 20 ng/mL recombinant human FGF-2 (Peprotech).

    Techniques: Expressing, Generated, In Vitro, In Vivo

    Application of PF6 to demonstrate that adult hippocampal stem/progenitor cells (AHPs) produce H 2 O 2 upon FGF-2 stimulation (a) After FGF-2 starvation, AHPs were loaded with 5 μM PF6-AM for 30 minutes, stimulated with 20 ng/mL FGF-2 or media for 30 minutes, and then imaged. For DPI treatment, cells were preincubated in media containing 5 μM DPI before FGF-2 stimulation. (b) AHPs were transfected with either Catalase or control vector and treated as in (a). (c) AHPs were transfected with either Nox2-shRNA or control vector and treated as in (a). Brightfield images are shown for each representative image with a 50 μm scale bar.

    Journal: Nature chemical biology

    Article Title: Nox2 redox signaling maintains essential cell populations in the brain

    doi: 10.1038/nchembio.497

    Figure Lengend Snippet: Application of PF6 to demonstrate that adult hippocampal stem/progenitor cells (AHPs) produce H 2 O 2 upon FGF-2 stimulation (a) After FGF-2 starvation, AHPs were loaded with 5 μM PF6-AM for 30 minutes, stimulated with 20 ng/mL FGF-2 or media for 30 minutes, and then imaged. For DPI treatment, cells were preincubated in media containing 5 μM DPI before FGF-2 stimulation. (b) AHPs were transfected with either Catalase or control vector and treated as in (a). (c) AHPs were transfected with either Nox2-shRNA or control vector and treated as in (a). Brightfield images are shown for each representative image with a 50 μm scale bar.

    Article Snippet: Cell Culture AHPs were cultured on tissue culture polystyrene coated with poly-ornithine and 5 μg/mL of laminin (Invitrogen) and grown in Dulbecco's modified Eagle medium (DMEM)/F-12 (1:1) high-glucose medium (Invitrogen) containing N-2 supplement (Invitrogen) and 20 ng/mL recombinant human FGF-2 (Peprotech).

    Techniques: Transfection, Plasmid Preparation, shRNA

    Cellular redox status affects AHP growth signaling (a) After FGF-2 starvation, AHPs were stimulated with vehicle control (buffer), 20 ng/mL FGF-2, 300, 500, or 1000 μM H 2 O 2 for 30 min. (b) AHPs transfected with either Catalase or a control vector. After FGF-2 starvation, AHPs were stimulated with 20 ng/mL and lysed at the given time points. (c) After FGF-2 starvation, AHPs were incubated with NAC, DPI, or vehicle control (DMSO) for 40 minutes, then stimulated with 20 ng/mL FGF-2 and lysed at the given time points. (d) Nox2 mRNA detection in AHPs measured by RT-PCR. (e) Nox2 expression of AHP whole cell extracts transfected with either Nox2 shRNA or an empty vector as measured by western blot analysis using either a mouse monoclonal (m) or a rabbit polyclonal (r) Nox2 antibody, followed by stripping and reprobing for actin as a loading control. An arrow shows the band in the Nox2 monoclonal antibody blot that matches the band in the Nox2 polyclonal blot, which corresponds to the molecular weight of Nox2. (f) AHPs transfected with either Nox2 shRNA or the empty vector. (g) AHPs transfected with either Nox2 shRNA or Nox3 shRNA. After 12 hour FGF-2 starvation, AHPs were stimulated with 20 ng/mL and lysed at the given time points. phospho-Akt, phospho-ERK, or mouse monoclonal Nox2 were measured by western blot analysis of whole cell extracts, and blots were stripped and reprobed for total protein or actin as loading controls.

    Journal: Nature chemical biology

    Article Title: Nox2 redox signaling maintains essential cell populations in the brain

    doi: 10.1038/nchembio.497

    Figure Lengend Snippet: Cellular redox status affects AHP growth signaling (a) After FGF-2 starvation, AHPs were stimulated with vehicle control (buffer), 20 ng/mL FGF-2, 300, 500, or 1000 μM H 2 O 2 for 30 min. (b) AHPs transfected with either Catalase or a control vector. After FGF-2 starvation, AHPs were stimulated with 20 ng/mL and lysed at the given time points. (c) After FGF-2 starvation, AHPs were incubated with NAC, DPI, or vehicle control (DMSO) for 40 minutes, then stimulated with 20 ng/mL FGF-2 and lysed at the given time points. (d) Nox2 mRNA detection in AHPs measured by RT-PCR. (e) Nox2 expression of AHP whole cell extracts transfected with either Nox2 shRNA or an empty vector as measured by western blot analysis using either a mouse monoclonal (m) or a rabbit polyclonal (r) Nox2 antibody, followed by stripping and reprobing for actin as a loading control. An arrow shows the band in the Nox2 monoclonal antibody blot that matches the band in the Nox2 polyclonal blot, which corresponds to the molecular weight of Nox2. (f) AHPs transfected with either Nox2 shRNA or the empty vector. (g) AHPs transfected with either Nox2 shRNA or Nox3 shRNA. After 12 hour FGF-2 starvation, AHPs were stimulated with 20 ng/mL and lysed at the given time points. phospho-Akt, phospho-ERK, or mouse monoclonal Nox2 were measured by western blot analysis of whole cell extracts, and blots were stripped and reprobed for total protein or actin as loading controls.

    Article Snippet: Cell Culture AHPs were cultured on tissue culture polystyrene coated with poly-ornithine and 5 μg/mL of laminin (Invitrogen) and grown in Dulbecco's modified Eagle medium (DMEM)/F-12 (1:1) high-glucose medium (Invitrogen) containing N-2 supplement (Invitrogen) and 20 ng/mL recombinant human FGF-2 (Peprotech).

    Techniques: Transfection, Plasmid Preparation, Incubation, Reverse Transcription Polymerase Chain Reaction, Expressing, shRNA, Western Blot, Stripping Membranes, Molecular Weight

    Basic fibroblast growth factor ( bFGF) knockdown inhibits paracrine signaling‐mediated activation of endothelial cell proliferation and tube assembly in HDAC 1‐depleted CMC s. A, Immunoblots evaluating the expression of bFGF in total protein extracts derived from sh RNA ‐transduced CMC s (n=4). B, Densitometric analysis of bFGF immunoblots (expression relative to β‐actin). Values are mean± SEM (n=4). Western blot data were log base 10 (y=log 10 y) transformed and analyzed by unpaired, 1‐way ANOVA . P values were calculated using the post hoc Holm–Sidak multiple comparison test. C, HAEC growth was assessed following their propagation in CM from all sh RNA ‐transduced CMC groups (untransduced, sh NT , sh HDAC 1, shb FGF , and sh HDAC 1+shb FGF ). Values are mean± SEM (n=11). Growth data were analyzed by 1‐way ANOVA and P values determined using the post hoc Dunnett multiple comparison test. D, Representative fluorescent microscopy images (scale=1000 μm) and (E) graph enumerating HUVEC tube formation in response to incubation with CM from untransduced, sh NT , sh HDAC 1, shb FGF , or sh HDAC 1+shb FGF transduced CMC s. HUVEC base (−Ctrl), CMC base (−Ctrl), and HUVEC base+inducer (low serum growth supplement [ LSGS ]; +Ctrl) controls are included. Values are mean± SEM (n=8 for control and experimental groups). Tube formation data were analyzed using 1‐way ANOVA and P values determined using the post hoc Dunnett multiple comparison test. All experiments utilized 1:3 sh RNA viral titer dilutions. CM indicates conditioned medium; CMC , cardiac mesenchymal stromal cell; HAEC , human aortic endothelial cell; HDAC 1, histone deacetylase 1; HUVEC , human umbilical vein endothelial cell; shb FGF , short hairpin RNA basic fibroblast growth factor; sh HDAC 1, short hairpin RNA ‐histone deacetylase 1; sh NT , short hairpin RNA nontarget; UT , untransduced.

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Histone Deacetylase 1 Depletion Activates Human Cardiac Mesenchymal Stromal Cell Proangiogenic Paracrine Signaling Through a Mechanism Requiring Enhanced Basic Fibroblast Growth Factor Synthesis and Secretion

    doi: 10.1161/JAHA.117.006183

    Figure Lengend Snippet: Basic fibroblast growth factor ( bFGF) knockdown inhibits paracrine signaling‐mediated activation of endothelial cell proliferation and tube assembly in HDAC 1‐depleted CMC s. A, Immunoblots evaluating the expression of bFGF in total protein extracts derived from sh RNA ‐transduced CMC s (n=4). B, Densitometric analysis of bFGF immunoblots (expression relative to β‐actin). Values are mean± SEM (n=4). Western blot data were log base 10 (y=log 10 y) transformed and analyzed by unpaired, 1‐way ANOVA . P values were calculated using the post hoc Holm–Sidak multiple comparison test. C, HAEC growth was assessed following their propagation in CM from all sh RNA ‐transduced CMC groups (untransduced, sh NT , sh HDAC 1, shb FGF , and sh HDAC 1+shb FGF ). Values are mean± SEM (n=11). Growth data were analyzed by 1‐way ANOVA and P values determined using the post hoc Dunnett multiple comparison test. D, Representative fluorescent microscopy images (scale=1000 μm) and (E) graph enumerating HUVEC tube formation in response to incubation with CM from untransduced, sh NT , sh HDAC 1, shb FGF , or sh HDAC 1+shb FGF transduced CMC s. HUVEC base (−Ctrl), CMC base (−Ctrl), and HUVEC base+inducer (low serum growth supplement [ LSGS ]; +Ctrl) controls are included. Values are mean± SEM (n=8 for control and experimental groups). Tube formation data were analyzed using 1‐way ANOVA and P values determined using the post hoc Dunnett multiple comparison test. All experiments utilized 1:3 sh RNA viral titer dilutions. CM indicates conditioned medium; CMC , cardiac mesenchymal stromal cell; HAEC , human aortic endothelial cell; HDAC 1, histone deacetylase 1; HUVEC , human umbilical vein endothelial cell; shb FGF , short hairpin RNA basic fibroblast growth factor; sh HDAC 1, short hairpin RNA ‐histone deacetylase 1; sh NT , short hairpin RNA nontarget; UT , untransduced.

    Article Snippet: Cells, Cell Culture, and Treatment Primary patient‐derived CMCs were propagated in Ham's F12 medium (Gibco, Grand Island, NY) supplemented with 10% FBS (Seradigm, Radnor, PA), 20 ng/mL of recombinant human basic fibroblast growth factor (bFGF; PeproTech, Rocky Hill, NJ), 0.2 mmol/L of l ‐glutamine (Gibco), 0.005 U/mL of human erythropoietin (Invitrogen, Carlsbad, CA), and 100 U/mL of penicillin/streptomycin (Gibco).

    Techniques: Activation Assay, Western Blot, Expressing, Derivative Assay, Transformation Assay, Microscopy, Incubation, Histone Deacetylase Assay, shRNA

    HDAC 1‐depletion stimulates basic fibroblast growth factor (bFGF) expression in human CMC s. A, q PCR assays evaluating the expression of known trophic factors involved in cell‐mediated cardiac repair in sh HDAC 1, sh NT , or untransduced CMC s. Values are mean± SEM (n=4). qPCR data were log base 10 (y=log 10 y) transformed and analyzed by 2‐way ANOVA . P values were calculated using the post hoc Bonferroni multiple comparison test. B, Representative immunoblot evaluating bFGF expression in total protein isolates derived from sh HDAC 1, sh NT , or untransduced CMC s (n=4). C, Densitometric quantification of bFGF immunoblots (expression relative to β‐actin). Values are mean± SEM (n=4). Western blot data were log base 10 (y=log 10 y) transformed and analyzed by unpaired, 1‐way ANOVA . P values were calculated using the post hoc Bonferroni multiple comparison test. All experiments utilized 1:3 sh RNA viral titer dilutions. HDAC 1 indicates histone deacetylase 1; sh HDAC 1, short hairpin RNA ‐histone deacetylase 1; sh NT , short hairpin RNA ‐non target; UT , untransduced.

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Histone Deacetylase 1 Depletion Activates Human Cardiac Mesenchymal Stromal Cell Proangiogenic Paracrine Signaling Through a Mechanism Requiring Enhanced Basic Fibroblast Growth Factor Synthesis and Secretion

    doi: 10.1161/JAHA.117.006183

    Figure Lengend Snippet: HDAC 1‐depletion stimulates basic fibroblast growth factor (bFGF) expression in human CMC s. A, q PCR assays evaluating the expression of known trophic factors involved in cell‐mediated cardiac repair in sh HDAC 1, sh NT , or untransduced CMC s. Values are mean± SEM (n=4). qPCR data were log base 10 (y=log 10 y) transformed and analyzed by 2‐way ANOVA . P values were calculated using the post hoc Bonferroni multiple comparison test. B, Representative immunoblot evaluating bFGF expression in total protein isolates derived from sh HDAC 1, sh NT , or untransduced CMC s (n=4). C, Densitometric quantification of bFGF immunoblots (expression relative to β‐actin). Values are mean± SEM (n=4). Western blot data were log base 10 (y=log 10 y) transformed and analyzed by unpaired, 1‐way ANOVA . P values were calculated using the post hoc Bonferroni multiple comparison test. All experiments utilized 1:3 sh RNA viral titer dilutions. HDAC 1 indicates histone deacetylase 1; sh HDAC 1, short hairpin RNA ‐histone deacetylase 1; sh NT , short hairpin RNA ‐non target; UT , untransduced.

    Article Snippet: Cells, Cell Culture, and Treatment Primary patient‐derived CMCs were propagated in Ham's F12 medium (Gibco, Grand Island, NY) supplemented with 10% FBS (Seradigm, Radnor, PA), 20 ng/mL of recombinant human basic fibroblast growth factor (bFGF; PeproTech, Rocky Hill, NJ), 0.2 mmol/L of l ‐glutamine (Gibco), 0.005 U/mL of human erythropoietin (Invitrogen, Carlsbad, CA), and 100 U/mL of penicillin/streptomycin (Gibco).

    Techniques: Expressing, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Transformation Assay, Derivative Assay, Western Blot, Histone Deacetylase Assay, shRNA

    ENC spheroid culture. a) Protocol (days 12–15) for ENC spheroid formation. NB+N2+B27; NB/N2/B27, Neurobasal medium with N2 and B27 supplement; FGF2, Recombinant Human FGF Basic; CHIR, CHIR 99021. b) Phase contrast and SOX10::GFP reporter line GFP expression of 3D spheroids on day 14. Scale bar = 200 μm.

    Journal: Nature protocols

    Article Title: DERIVATION OF ENTERIC NEURON LINEAGES FROM HUMAN PLURIPOTENT STEM CELLS

    doi: 10.1038/s41596-019-0141-y

    Figure Lengend Snippet: ENC spheroid culture. a) Protocol (days 12–15) for ENC spheroid formation. NB+N2+B27; NB/N2/B27, Neurobasal medium with N2 and B27 supplement; FGF2, Recombinant Human FGF Basic; CHIR, CHIR 99021. b) Phase contrast and SOX10::GFP reporter line GFP expression of 3D spheroids on day 14. Scale bar = 200 μm.

    Article Snippet: FGF2, Recombinant Human FGF Basic (R & D Systems #233-FB) Critical: Stock aliquots should be stored at −80 °C.

    Techniques: Recombinant, Expressing

    Intronic precursor of miR-4673 in notch-1 codes an active enhancer. a, Structural analysis shows improved thermodynamic stability of the RNA hairpin in primate lineage culminating in the structural maturation of miR-4673 hairpin in Hominins . b, Nucleosome-favouring dinucleotide usage map of human notch-1 intron 4. c, Transmission electron micrograph of a nucleosome formed by NPS miR (left). Gel retardation (middle) and restriction enzyme digestion assays (right) provided further confirmation for nucleosome formation by NPS miR . d , The temporal profile of enhancer-RNAs (eRNAs) that originates from Notch-1 intron-4 (full blots are provided in supplementary file 10). Vertical lines show the location of eRNAs with reference to the dinucleotide usage map. In order to access the eRNA profile, cells were synchronised at G0 and released into G1 by application of a single pulse of FGF-2. Note the temporal stability of 3′-eRNA from a region that corresponds to NPS miR (see supplementary file 11). This region is positioned at 3′-terminus of a stable RNA palindrome formed by two Alu elements. e, Histogram shows cumulative distribution of TCF3/4 and TFAP2A/B/C cis -motifs in the intron 4 of human notch-1. At the bottom, Levenshtein distance (L.E.D) as a measure of intronic change is aligned to the enhancer map. Sequences that belong to AluJb and AluYa1 insertions in all Simians are in grey. Species-specific transpositional events are marked in LED heat maps.

    Journal: bioRxiv

    Article Title: Genomic Competition for Noise Reduction Shaped Evolutionary Landscape of Mir-4673

    doi: 10.1101/788984

    Figure Lengend Snippet: Intronic precursor of miR-4673 in notch-1 codes an active enhancer. a, Structural analysis shows improved thermodynamic stability of the RNA hairpin in primate lineage culminating in the structural maturation of miR-4673 hairpin in Hominins . b, Nucleosome-favouring dinucleotide usage map of human notch-1 intron 4. c, Transmission electron micrograph of a nucleosome formed by NPS miR (left). Gel retardation (middle) and restriction enzyme digestion assays (right) provided further confirmation for nucleosome formation by NPS miR . d , The temporal profile of enhancer-RNAs (eRNAs) that originates from Notch-1 intron-4 (full blots are provided in supplementary file 10). Vertical lines show the location of eRNAs with reference to the dinucleotide usage map. In order to access the eRNA profile, cells were synchronised at G0 and released into G1 by application of a single pulse of FGF-2. Note the temporal stability of 3′-eRNA from a region that corresponds to NPS miR (see supplementary file 11). This region is positioned at 3′-terminus of a stable RNA palindrome formed by two Alu elements. e, Histogram shows cumulative distribution of TCF3/4 and TFAP2A/B/C cis -motifs in the intron 4 of human notch-1. At the bottom, Levenshtein distance (L.E.D) as a measure of intronic change is aligned to the enhancer map. Sequences that belong to AluJb and AluYa1 insertions in all Simians are in grey. Species-specific transpositional events are marked in LED heat maps.

    Article Snippet: DMEM/F12 supplemented with 10% fetal calf serum (FCS), recombinant human FGF-2 (20 ng/ml, R & D Systems) and Antibiotic-Antimycotic® (100X, Life Technologies) was used for culturing the cells.

    Techniques: Transmission Assay, Electrophoretic Mobility Shift Assay