fibroblast growth factor fgf Search Results


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  • 99
    Thermo Fisher fgf 2
    Shh induces tectal NSC proliferation. Nsp suspension cultures were immobilized in collagen type-I gels. Proliferation experiments were performed for 8 hours and included a BrdU (1 µg/ml) pulse for the last 2 hours in the presence of EGF and <t>FGF-2</t> (used at 1 ng/ml or 10 ng/ml); combined with either Shh (3,3 µg/ml) or Cyc (5 or 10 µM). ( A ) Representative images of double immunostaining for BrdU and the RGC marker Blbp after indicated treatments. Bar, 20 µm. ( B ) Quantification of the percentage of positive cells for Blbp over total numbers of cells, in Z-stacks of confocal microscope. Anti-Shh antibody 5E1 affects both endogenous Shh source as well as recombinant Shh effect resulting in less number of Blbp positive cells. ( C ) Quantification of the effect of endogenous Shh on the number of Blbp positive cells shows significant inhibition after Cyc treatment. ( D ) Histogram showing proliferation of nsps after exposures to treatments as indicated. The anti-Shh antibody 5E1 reduces significantly number of BrdU positive cells in the Blbp positive cell population. ( E ) Quantification of positive cells for BrdU over Blbp positive cells in Z-stacks of confocal microscope treated with Cyc. *, p
    Fgf 2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1044 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore fgf 2
    Inhibition of TGF-α signaling blocks tube formation and cell survival induced by TGF-β1 in collagen gels. (Top) 1G11 endothelial cells grown in collagen gels were cultured for 24 h in the presence of DMEM alone (basal) or 10 ng of TGF-β1/ml either alone or supplemented with 1 μM tyrphostin AG1478. After this time, cells were examined by phase-contrast microscopy. Magnifications: left, ×90; right, ×180. (Bottom) 1G11 endothelial cells grown in collagen gels were cultured for 24 h in the presence of DMEM either alone (lane B) or with 1 μM tyrphostin AG1478 (B+AG), in 10 ng of TGF-β1/ml either alone (TGFβ) or with 1 μM tyrphostin AG1478 (TGFβ+AG), in 50 ng of TGF-α/ml either alone (TGFα) or with tyrphostin AG1478 (TGFα+AG), or in 100 ng of <t>FGF-2/ml</t> either alone (FGF) or with 1 μM tyrphostin AG1478 (FGF+AG). Dead and living cells were counted as described in Materials and Methods. Results are averages of three different experiments.
    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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    Millipore bfgf
    ERK5 signaling regulates neurogenesis of SVZ-derived aNPCs in culture. (A) ERK5 signaling is necessary for promoting spontaneous neurogenesis. aNPCs were infected with non-specific shRNA control retroviral vector (shNS) or shRNA to ERK5 retrovirus (shERK5). Both retroviral vectors encode an eGFP marker protein under a bicistronic promoter [26] . One day after virus infection, cells were incubated in <t>EGF</t> and <t>bFGF-free</t> medium for 5 d to allow spontaneous differentiation. The percentage of GFP + cells that were also SOX2 + , PCNA + , or β-III Tubulin + was quantified. (B) Activation of endogenous ERK5 signaling is sufficient to promote neurogenesis. aNPCs were infected with control retroviral vector expressing eGFP only or expressing caMEK5-IRES-eGFP. One day after virus infection, cells were washed and then placed in fresh regular medium containing mitogenic EGF and bFGF for 5 d. (C) ERK5 knockdown does not affect glial differentiation. GFAP, a marker for astrocytes as well as SVZ stem cells; S100β, a marker for astrocytes; O4, an oligodendrocyte marker. (D) Effect of ERK5 activation on cells expressing GFAP, S100β, and O4. Over 200 virus-infected cells (GFP + ) from each sample were analyzed and quantified. Data are mean ± SEM from three independent experiments (n = 3). *, p
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    94
    Thermo Fisher fibroblast growth factor
    ERK5 signaling regulates neurogenesis of SVZ-derived aNPCs in culture. (A) ERK5 signaling is necessary for promoting spontaneous neurogenesis. aNPCs were infected with non-specific shRNA control retroviral vector (shNS) or shRNA to ERK5 retrovirus (shERK5). Both retroviral vectors encode an eGFP marker protein under a bicistronic promoter [26] . One day after virus infection, cells were incubated in <t>EGF</t> and <t>bFGF-free</t> medium for 5 d to allow spontaneous differentiation. The percentage of GFP + cells that were also SOX2 + , PCNA + , or β-III Tubulin + was quantified. (B) Activation of endogenous ERK5 signaling is sufficient to promote neurogenesis. aNPCs were infected with control retroviral vector expressing eGFP only or expressing caMEK5-IRES-eGFP. One day after virus infection, cells were washed and then placed in fresh regular medium containing mitogenic EGF and bFGF for 5 d. (C) ERK5 knockdown does not affect glial differentiation. GFAP, a marker for astrocytes as well as SVZ stem cells; S100β, a marker for astrocytes; O4, an oligodendrocyte marker. (D) Effect of ERK5 activation on cells expressing GFAP, S100β, and O4. Over 200 virus-infected cells (GFP + ) from each sample were analyzed and quantified. Data are mean ± SEM from three independent experiments (n = 3). *, p
    Fibroblast Growth Factor, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 889 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore fibroblast growth factor
    ERK5 signaling regulates neurogenesis of SVZ-derived aNPCs in culture. (A) ERK5 signaling is necessary for promoting spontaneous neurogenesis. aNPCs were infected with non-specific shRNA control retroviral vector (shNS) or shRNA to ERK5 retrovirus (shERK5). Both retroviral vectors encode an eGFP marker protein under a bicistronic promoter [26] . One day after virus infection, cells were incubated in <t>EGF</t> and <t>bFGF-free</t> medium for 5 d to allow spontaneous differentiation. The percentage of GFP + cells that were also SOX2 + , PCNA + , or β-III Tubulin + was quantified. (B) Activation of endogenous ERK5 signaling is sufficient to promote neurogenesis. aNPCs were infected with control retroviral vector expressing eGFP only or expressing caMEK5-IRES-eGFP. One day after virus infection, cells were washed and then placed in fresh regular medium containing mitogenic EGF and bFGF for 5 d. (C) ERK5 knockdown does not affect glial differentiation. GFAP, a marker for astrocytes as well as SVZ stem cells; S100β, a marker for astrocytes; O4, an oligodendrocyte marker. (D) Effect of ERK5 activation on cells expressing GFAP, S100β, and O4. Over 200 virus-infected cells (GFP + ) from each sample were analyzed and quantified. Data are mean ± SEM from three independent experiments (n = 3). *, p
    Fibroblast Growth Factor, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 978 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore basic fibroblast growth factor bfgf
    Inhibition of angiogenesis by ADI in vivo mouse Matrigel assay. Matrigel (0.4 ml) containing 50 ng ml −1 of <t>bFGF</t> and 60 U ml −1 of heparin in combination with or without 0.46 U ml −1 ADI was subcutaneously injected near the abdominal midline of the mice. ( A ) Histological analysis of Matrigel implants (for experimental procedures, see Materials and methods). Matrigel without ADI (A and B) showed formation of blood vessels with various sizes (arrows) forming in the Matrigel. Inside the vessel, red blood cells were observed (red colour in the vessel). However, ADI treatment clearly inhibited blood vessel formation (C and D). * Indicates connective tissues surrounding Matrigel implants. (A and C: haematoxylin–eosin staining, B and D: Masson-Trichrome staining). Original magnification x100. ( B ) Haemoglobin content in the Matrigel was measured with <t>Drabkin</t> reagent kit 525 to evaluate blood within the vessels formed 5 days after injection, calibrated against a known amount of haemoglobin in parallel. ADI potently inhibited growth factor-induced angiogenesis by 97%. Each value represents the mean±s.e.of five ADI-treated animals and seven per control group.
    Basic Fibroblast Growth Factor Bfgf, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 841 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech fibroblast growth factor 2
    TGFβ-driven Smad2/3 phosphorylation and stem cell chondrogenesis. hMSCs were cultured on  (A)  Tissue cultured plastic in chondrogenic media supplemented with transforming growth factor-β (TGFβ) and  (B)  composite cellulose:silk 75:25 substrates in stem cell expansion media supplemented with 10 ng/ml fibroblast growth factor 2, with and without the SB505124 inhibitor. Cells were cultured for 14 days after which gene analysis was performed. Gene expression levels were normalized to expression levels of housekeeping gene, GAPDH (shown in dotted line). Data points show mean ± SE. Statistical significance was assessed using a two-way ANOVA with Bonferroni  post-test . ** p
    Fibroblast Growth Factor 2, supplied by PeproTech, used in various techniques. Bioz Stars score: 97/100, based on 686 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher human basic fibroblast growth factor
    TGFβ-driven Smad2/3 phosphorylation and stem cell chondrogenesis. hMSCs were cultured on  (A)  Tissue cultured plastic in chondrogenic media supplemented with transforming growth factor-β (TGFβ) and  (B)  composite cellulose:silk 75:25 substrates in stem cell expansion media supplemented with 10 ng/ml fibroblast growth factor 2, with and without the SB505124 inhibitor. Cells were cultured for 14 days after which gene analysis was performed. Gene expression levels were normalized to expression levels of housekeeping gene, GAPDH (shown in dotted line). Data points show mean ± SE. Statistical significance was assessed using a two-way ANOVA with Bonferroni  post-test . ** p
    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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    Becton Dickinson basic fibroblast growth factor
    TGFβ-driven Smad2/3 phosphorylation and stem cell chondrogenesis. hMSCs were cultured on  (A)  Tissue cultured plastic in chondrogenic media supplemented with transforming growth factor-β (TGFβ) and  (B)  composite cellulose:silk 75:25 substrates in stem cell expansion media supplemented with 10 ng/ml fibroblast growth factor 2, with and without the SB505124 inhibitor. Cells were cultured for 14 days after which gene analysis was performed. Gene expression levels were normalized to expression levels of housekeeping gene, GAPDH (shown in dotted line). Data points show mean ± SE. Statistical significance was assessed using a two-way ANOVA with Bonferroni  post-test . ** p
    Basic Fibroblast Growth Factor, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 250 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega basic fibroblast growth factor
    TGFβ-driven Smad2/3 phosphorylation and stem cell chondrogenesis. hMSCs were cultured on  (A)  Tissue cultured plastic in chondrogenic media supplemented with transforming growth factor-β (TGFβ) and  (B)  composite cellulose:silk 75:25 substrates in stem cell expansion media supplemented with 10 ng/ml fibroblast growth factor 2, with and without the SB505124 inhibitor. Cells were cultured for 14 days after which gene analysis was performed. Gene expression levels were normalized to expression levels of housekeeping gene, GAPDH (shown in dotted line). Data points show mean ± SE. Statistical significance was assessed using a two-way ANOVA with Bonferroni  post-test . ** p
    Basic Fibroblast Growth Factor, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 344 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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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    FUJIFILM 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.
    Basic Fibroblast Growth Factor, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 93/100, based on 274 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher recombinant human basic fibroblast growth factor
    <t>Growth</t> <t>factor</t> 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 <t>recombinant</t> <t>human</t> <t>basic</t> <t>fibroblast</t> 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.
    Recombinant Human Basic Fibroblast Growth Factor, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 235 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    FUJIFILM recombinant human basic fibroblast growth factor
    <t>Growth</t> <t>factor</t> 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 <t>recombinant</t> <t>human</t> <t>basic</t> <t>fibroblast</t> 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.
    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
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    PromoCell basic fibroblast growth factor
    <t>Growth</t> <t>factor</t> 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 <t>recombinant</t> <t>human</t> <t>basic</t> <t>fibroblast</t> 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.
    Basic Fibroblast Growth Factor, supplied by PromoCell, used in various techniques. Bioz Stars score: 92/100, based on 198 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
    <t>Growth</t> <t>factor</t> 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 <t>recombinant</t> <t>human</t> <t>basic</t> <t>fibroblast</t> 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.
    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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    Prospec basic fibroblast growth factor
    <t>Growth</t> <t>factor</t> 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 <t>recombinant</t> <t>human</t> <t>basic</t> <t>fibroblast</t> 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.
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    Thermo Fisher fgf basic aa 10 155 recombinant human protein
    <t>Growth</t> <t>factor</t> 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 <t>recombinant</t> <t>human</t> <t>basic</t> <t>fibroblast</t> 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.
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    Becton Dickinson fibroblast growth factor
    <t>Growth</t> <t>factor</t> 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 <t>recombinant</t> <t>human</t> <t>basic</t> <t>fibroblast</t> 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.
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    Millipore recombinant human basic fibroblast growth factor
    <t>Growth</t> <t>factor</t> 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 <t>recombinant</t> <t>human</t> <t>basic</t> <t>fibroblast</t> 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.
    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
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    Image Search Results


    Shh induces tectal NSC proliferation. Nsp suspension cultures were immobilized in collagen type-I gels. Proliferation experiments were performed for 8 hours and included a BrdU (1 µg/ml) pulse for the last 2 hours in the presence of EGF and FGF-2 (used at 1 ng/ml or 10 ng/ml); combined with either Shh (3,3 µg/ml) or Cyc (5 or 10 µM). ( A ) Representative images of double immunostaining for BrdU and the RGC marker Blbp after indicated treatments. Bar, 20 µm. ( B ) Quantification of the percentage of positive cells for Blbp over total numbers of cells, in Z-stacks of confocal microscope. Anti-Shh antibody 5E1 affects both endogenous Shh source as well as recombinant Shh effect resulting in less number of Blbp positive cells. ( C ) Quantification of the effect of endogenous Shh on the number of Blbp positive cells shows significant inhibition after Cyc treatment. ( D ) Histogram showing proliferation of nsps after exposures to treatments as indicated. The anti-Shh antibody 5E1 reduces significantly number of BrdU positive cells in the Blbp positive cell population. ( E ) Quantification of positive cells for BrdU over Blbp positive cells in Z-stacks of confocal microscope treated with Cyc. *, p

    Journal: PLoS ONE

    Article Title: Proliferation of Murine Midbrain Neural Stem Cells Depends upon an Endogenous Sonic Hedgehog (Shh) Source

    doi: 10.1371/journal.pone.0065818

    Figure Lengend Snippet: Shh induces tectal NSC proliferation. Nsp suspension cultures were immobilized in collagen type-I gels. Proliferation experiments were performed for 8 hours and included a BrdU (1 µg/ml) pulse for the last 2 hours in the presence of EGF and FGF-2 (used at 1 ng/ml or 10 ng/ml); combined with either Shh (3,3 µg/ml) or Cyc (5 or 10 µM). ( A ) Representative images of double immunostaining for BrdU and the RGC marker Blbp after indicated treatments. Bar, 20 µm. ( B ) Quantification of the percentage of positive cells for Blbp over total numbers of cells, in Z-stacks of confocal microscope. Anti-Shh antibody 5E1 affects both endogenous Shh source as well as recombinant Shh effect resulting in less number of Blbp positive cells. ( C ) Quantification of the effect of endogenous Shh on the number of Blbp positive cells shows significant inhibition after Cyc treatment. ( D ) Histogram showing proliferation of nsps after exposures to treatments as indicated. The anti-Shh antibody 5E1 reduces significantly number of BrdU positive cells in the Blbp positive cell population. ( E ) Quantification of positive cells for BrdU over Blbp positive cells in Z-stacks of confocal microscope treated with Cyc. *, p

    Article Snippet: Other treatments included Hh inhibitor Cyclopamine (Cyc) at 5 µM and 10 µM (Infinity Pharmaceuticals, Inc.), Hh agonist Purmorphamine (Pur) at 10 µM (Infinity Pharmaceuticals, Inc.), EGF 1 and 10 ng/ml (human recombinant, Invitrogen) and/or FGF-2 at 1 and 10 ng/ml (Invitrogen).

    Techniques: Double Immunostaining, Marker, Microscopy, Recombinant, Inhibition

    Shh promotes proliferation and maintenance of RGCs. Differentiation experiments were performed during 14 days without addition of EGF or FGF-2. ( A ) Representative images of immunostaining for Blbp after control or Shh treatment. Close-up view of a RGC stained for Blbp treated with Shh. Bar, 20 µm. ( B ) Western blot and densitometry analysis for Blbp expression show higher levels in Shh treated cultures. Western Blot analysis of Sox2 ( C ), and Nestin ( D ) levels after 7 days of treatment with Shh indicate an increase of neural progenitors. ( E ) Western blot of Cyclin D1, a read-out response to Shh pathway activation, indicates an increased proliferation even in absence of other additional growth factors. *, p

    Journal: PLoS ONE

    Article Title: Proliferation of Murine Midbrain Neural Stem Cells Depends upon an Endogenous Sonic Hedgehog (Shh) Source

    doi: 10.1371/journal.pone.0065818

    Figure Lengend Snippet: Shh promotes proliferation and maintenance of RGCs. Differentiation experiments were performed during 14 days without addition of EGF or FGF-2. ( A ) Representative images of immunostaining for Blbp after control or Shh treatment. Close-up view of a RGC stained for Blbp treated with Shh. Bar, 20 µm. ( B ) Western blot and densitometry analysis for Blbp expression show higher levels in Shh treated cultures. Western Blot analysis of Sox2 ( C ), and Nestin ( D ) levels after 7 days of treatment with Shh indicate an increase of neural progenitors. ( E ) Western blot of Cyclin D1, a read-out response to Shh pathway activation, indicates an increased proliferation even in absence of other additional growth factors. *, p

    Article Snippet: Other treatments included Hh inhibitor Cyclopamine (Cyc) at 5 µM and 10 µM (Infinity Pharmaceuticals, Inc.), Hh agonist Purmorphamine (Pur) at 10 µM (Infinity Pharmaceuticals, Inc.), EGF 1 and 10 ng/ml (human recombinant, Invitrogen) and/or FGF-2 at 1 and 10 ng/ml (Invitrogen).

    Techniques: Immunostaining, Staining, Western Blot, Expressing, Activation Assay

    Schematic representation of a conventional CR system and the i-CR system. A. Pathologically confirmed surgical tumor tissues were collected and dissociated for in vitro drug treatment and analysis. Biopsy tissues were first inoculated into mice, and the PDX tumor tissues were collected and subjected to the same analytical process as surgical tumors. B. The major difference between the i-CR system and the conventional CR system is the composition of the cell-culture medium. Here, the selective medium was complete medium lacking Wnt 3A, R-spondin-1, and Noggin. Y-27632, ROCK1 inhibitor; FBS, fetal bovine serum; DMEM/F-12, Dulbecco’s modified Eagle’s medium/Ham’s F-12 medium; bFGF, basic fibroblast growth factor; EGF, epidermal growth factor.

    Journal: American Journal of Cancer Research

    Article Title: Conditionally reprogrammed colorectal cancer cells combined with mouse avatars identify synergy between EGFR and MEK or CDK4/6 inhibitors

    doi:

    Figure Lengend Snippet: Schematic representation of a conventional CR system and the i-CR system. A. Pathologically confirmed surgical tumor tissues were collected and dissociated for in vitro drug treatment and analysis. Biopsy tissues were first inoculated into mice, and the PDX tumor tissues were collected and subjected to the same analytical process as surgical tumors. B. The major difference between the i-CR system and the conventional CR system is the composition of the cell-culture medium. Here, the selective medium was complete medium lacking Wnt 3A, R-spondin-1, and Noggin. Y-27632, ROCK1 inhibitor; FBS, fetal bovine serum; DMEM/F-12, Dulbecco’s modified Eagle’s medium/Ham’s F-12 medium; bFGF, basic fibroblast growth factor; EGF, epidermal growth factor.

    Article Snippet: The complete medium consisted of DMEM/F-12 basal medium, 2% fetal bovine serum (FBS), 10 ng/mL human epithelial growth factor (EGF) (Thermo Fisher), 10 μM Y-27632 (Selleckchem), 10 ng/mL basic fibroblast growth factor (bFGF) (Thermo Fisher), 10 mM nicotinamide (Sigma), 1X insulin-transferrin-selenium (Thermo Fisher), 1X non-essential amino acids (Thermo Fisher), 25 ng/mL mouse Wnt3a (Peprotech), 500 ng/mL human R-spondin-1 (Peprotech), 100 ng/mL Noggin, and 100 μg/mL Primocin (Vivogen).

    Techniques: In Vitro, Mouse Assay, Cell Culture, Modification

    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

    Inhibition of TGF-α signaling blocks tube formation and cell survival induced by TGF-β1 in collagen gels. (Top) 1G11 endothelial cells grown in collagen gels were cultured for 24 h in the presence of DMEM alone (basal) or 10 ng of TGF-β1/ml either alone or supplemented with 1 μM tyrphostin AG1478. After this time, cells were examined by phase-contrast microscopy. Magnifications: left, ×90; right, ×180. (Bottom) 1G11 endothelial cells grown in collagen gels were cultured for 24 h in the presence of DMEM either alone (lane B) or with 1 μM tyrphostin AG1478 (B+AG), in 10 ng of TGF-β1/ml either alone (TGFβ) or with 1 μM tyrphostin AG1478 (TGFβ+AG), in 50 ng of TGF-α/ml either alone (TGFα) or with tyrphostin AG1478 (TGFα+AG), or in 100 ng of FGF-2/ml either alone (FGF) or with 1 μM tyrphostin AG1478 (FGF+AG). Dead and living cells were counted as described in Materials and Methods. Results are averages of three different experiments.

    Journal: Molecular and Cellular Biology

    Article Title: Transforming Growth Factor ?1 (TGF-?1) Promotes Endothelial Cell Survival during In Vitro Angiogenesis via an Autocrine Mechanism Implicating TGF-? Signaling

    doi: 10.1128/MCB.21.21.7218-7230.2001

    Figure Lengend Snippet: Inhibition of TGF-α signaling blocks tube formation and cell survival induced by TGF-β1 in collagen gels. (Top) 1G11 endothelial cells grown in collagen gels were cultured for 24 h in the presence of DMEM alone (basal) or 10 ng of TGF-β1/ml either alone or supplemented with 1 μM tyrphostin AG1478. After this time, cells were examined by phase-contrast microscopy. Magnifications: left, ×90; right, ×180. (Bottom) 1G11 endothelial cells grown in collagen gels were cultured for 24 h in the presence of DMEM either alone (lane B) or with 1 μM tyrphostin AG1478 (B+AG), in 10 ng of TGF-β1/ml either alone (TGFβ) or with 1 μM tyrphostin AG1478 (TGFβ+AG), in 50 ng of TGF-α/ml either alone (TGFα) or with tyrphostin AG1478 (TGFα+AG), or in 100 ng of FGF-2/ml either alone (FGF) or with 1 μM tyrphostin AG1478 (FGF+AG). Dead and living cells were counted as described in Materials and Methods. Results are averages of three different experiments.

    Article Snippet: Human umbilical vein endothelial cells (HUVEC) were obtained as previously described ( ) and were cultivated in SFM (Gibco-BRL) supplemented with 20% FCS and 100 μg of heparin, 20 ng of FGF-2, and 10 ng of EGF (Sigma)/ml.

    Techniques: Inhibition, Cell Culture, Microscopy

    Stimulation with TGF-β1 causes EGF receptor activation. (A) Quiescent 1G11 cells were stimulated or not (lane B) for the indicated times with 10 ng of TGF-β1/ml and for 10 min with 25 ng of FGF-2/ml, 10 ng of EGF/ml, or 10 ng of PDGF-BB/ml. Cells were lysed, and proteins were incubated with Sepharose-WGL for 1 h. After being washed, the final pellet was resuspended in Laemmli sample buffer and loaded on a SDS–7.5% polyacrylamide gel. Phosphotyrosine-containing proteins were immunodetected by using a specific antibody. Arrow, phosphotyrosine-containing protein that appeared after TGF-β1 treatment. A representative Western blot is shown. (B) Cells were treated as for panel A and lysed, and the EGF receptor (EGFR) was immunoprecipitated (IP) by incubation with a specific anti-EGF receptor antibody preadsorbed to protein A-Sepharose beads. After being washed the pellet was treated as for panel A. WB, Western blot.

    Journal: Molecular and Cellular Biology

    Article Title: Transforming Growth Factor ?1 (TGF-?1) Promotes Endothelial Cell Survival during In Vitro Angiogenesis via an Autocrine Mechanism Implicating TGF-? Signaling

    doi: 10.1128/MCB.21.21.7218-7230.2001

    Figure Lengend Snippet: Stimulation with TGF-β1 causes EGF receptor activation. (A) Quiescent 1G11 cells were stimulated or not (lane B) for the indicated times with 10 ng of TGF-β1/ml and for 10 min with 25 ng of FGF-2/ml, 10 ng of EGF/ml, or 10 ng of PDGF-BB/ml. Cells were lysed, and proteins were incubated with Sepharose-WGL for 1 h. After being washed, the final pellet was resuspended in Laemmli sample buffer and loaded on a SDS–7.5% polyacrylamide gel. Phosphotyrosine-containing proteins were immunodetected by using a specific antibody. Arrow, phosphotyrosine-containing protein that appeared after TGF-β1 treatment. A representative Western blot is shown. (B) Cells were treated as for panel A and lysed, and the EGF receptor (EGFR) was immunoprecipitated (IP) by incubation with a specific anti-EGF receptor antibody preadsorbed to protein A-Sepharose beads. After being washed the pellet was treated as for panel A. WB, Western blot.

    Article Snippet: Human umbilical vein endothelial cells (HUVEC) were obtained as previously described ( ) and were cultivated in SFM (Gibco-BRL) supplemented with 20% FCS and 100 μg of heparin, 20 ng of FGF-2, and 10 ng of EGF (Sigma)/ml.

    Techniques: Activation Assay, Incubation, Western Blot, Immunoprecipitation

    ERK5 signaling regulates neurogenesis of SVZ-derived aNPCs in culture. (A) ERK5 signaling is necessary for promoting spontaneous neurogenesis. aNPCs were infected with non-specific shRNA control retroviral vector (shNS) or shRNA to ERK5 retrovirus (shERK5). Both retroviral vectors encode an eGFP marker protein under a bicistronic promoter [26] . One day after virus infection, cells were incubated in EGF and bFGF-free medium for 5 d to allow spontaneous differentiation. The percentage of GFP + cells that were also SOX2 + , PCNA + , or β-III Tubulin + was quantified. (B) Activation of endogenous ERK5 signaling is sufficient to promote neurogenesis. aNPCs were infected with control retroviral vector expressing eGFP only or expressing caMEK5-IRES-eGFP. One day after virus infection, cells were washed and then placed in fresh regular medium containing mitogenic EGF and bFGF for 5 d. (C) ERK5 knockdown does not affect glial differentiation. GFAP, a marker for astrocytes as well as SVZ stem cells; S100β, a marker for astrocytes; O4, an oligodendrocyte marker. (D) Effect of ERK5 activation on cells expressing GFAP, S100β, and O4. Over 200 virus-infected cells (GFP + ) from each sample were analyzed and quantified. Data are mean ± SEM from three independent experiments (n = 3). *, p

    Journal: PLoS ONE

    Article Title: Targeted Deletion of the ERK5 MAP Kinase Impairs Neuronal Differentiation, Migration, and Survival during Adult Neurogenesis in the Olfactory Bulb

    doi: 10.1371/journal.pone.0061948

    Figure Lengend Snippet: ERK5 signaling regulates neurogenesis of SVZ-derived aNPCs in culture. (A) ERK5 signaling is necessary for promoting spontaneous neurogenesis. aNPCs were infected with non-specific shRNA control retroviral vector (shNS) or shRNA to ERK5 retrovirus (shERK5). Both retroviral vectors encode an eGFP marker protein under a bicistronic promoter [26] . One day after virus infection, cells were incubated in EGF and bFGF-free medium for 5 d to allow spontaneous differentiation. The percentage of GFP + cells that were also SOX2 + , PCNA + , or β-III Tubulin + was quantified. (B) Activation of endogenous ERK5 signaling is sufficient to promote neurogenesis. aNPCs were infected with control retroviral vector expressing eGFP only or expressing caMEK5-IRES-eGFP. One day after virus infection, cells were washed and then placed in fresh regular medium containing mitogenic EGF and bFGF for 5 d. (C) ERK5 knockdown does not affect glial differentiation. GFAP, a marker for astrocytes as well as SVZ stem cells; S100β, a marker for astrocytes; O4, an oligodendrocyte marker. (D) Effect of ERK5 activation on cells expressing GFAP, S100β, and O4. Over 200 virus-infected cells (GFP + ) from each sample were analyzed and quantified. Data are mean ± SEM from three independent experiments (n = 3). *, p

    Article Snippet: Tissue samples were then spun down and resuspended in serum-free culture media consisting of DMEM/F12 (Invitrogen), 1× N2 supplement (Invitrogen), 1× B27 supplement without retinoic acid (Invitrogen), 100 U/mL penicillin/streptomycin (Invitrogen), 2 mM L-glutamine (Invitrogen), 2 µg/mL heparin (Sigma), 20 ng/mL EGF (EMD Chemicals), and 10 ng/mL bFGF (Millipore).

    Techniques: Derivative Assay, Infection, shRNA, Plasmid Preparation, Marker, Incubation, Activation Assay, Expressing

    Inhibition of angiogenesis by ADI in vivo mouse Matrigel assay. Matrigel (0.4 ml) containing 50 ng ml −1 of bFGF and 60 U ml −1 of heparin in combination with or without 0.46 U ml −1 ADI was subcutaneously injected near the abdominal midline of the mice. ( A ) Histological analysis of Matrigel implants (for experimental procedures, see Materials and methods). Matrigel without ADI (A and B) showed formation of blood vessels with various sizes (arrows) forming in the Matrigel. Inside the vessel, red blood cells were observed (red colour in the vessel). However, ADI treatment clearly inhibited blood vessel formation (C and D). * Indicates connective tissues surrounding Matrigel implants. (A and C: haematoxylin–eosin staining, B and D: Masson-Trichrome staining). Original magnification x100. ( B ) Haemoglobin content in the Matrigel was measured with Drabkin reagent kit 525 to evaluate blood within the vessels formed 5 days after injection, calibrated against a known amount of haemoglobin in parallel. ADI potently inhibited growth factor-induced angiogenesis by 97%. Each value represents the mean±s.e.of five ADI-treated animals and seven per control group.

    Journal: British Journal of Cancer

    Article Title: Arginine deiminase: a potential inhibitor of angiogenesis and tumour growth

    doi: 10.1038/sj.bjc.6601181

    Figure Lengend Snippet: Inhibition of angiogenesis by ADI in vivo mouse Matrigel assay. Matrigel (0.4 ml) containing 50 ng ml −1 of bFGF and 60 U ml −1 of heparin in combination with or without 0.46 U ml −1 ADI was subcutaneously injected near the abdominal midline of the mice. ( A ) Histological analysis of Matrigel implants (for experimental procedures, see Materials and methods). Matrigel without ADI (A and B) showed formation of blood vessels with various sizes (arrows) forming in the Matrigel. Inside the vessel, red blood cells were observed (red colour in the vessel). However, ADI treatment clearly inhibited blood vessel formation (C and D). * Indicates connective tissues surrounding Matrigel implants. (A and C: haematoxylin–eosin staining, B and D: Masson-Trichrome staining). Original magnification x100. ( B ) Haemoglobin content in the Matrigel was measured with Drabkin reagent kit 525 to evaluate blood within the vessels formed 5 days after injection, calibrated against a known amount of haemoglobin in parallel. ADI potently inhibited growth factor-induced angiogenesis by 97%. Each value represents the mean±s.e.of five ADI-treated animals and seven per control group.

    Article Snippet: Basic fibroblast growth factor (bFGF), endothelial cell growth supplement, heparin, and Drabkin reagent kit 525 were from Sigma (St Louis, MO, USA).

    Techniques: Inhibition, In Vivo, Matrigel Assay, Injection, Mouse Assay, Staining

    TGFβ-driven Smad2/3 phosphorylation and stem cell chondrogenesis. hMSCs were cultured on  (A)  Tissue cultured plastic in chondrogenic media supplemented with transforming growth factor-β (TGFβ) and  (B)  composite cellulose:silk 75:25 substrates in stem cell expansion media supplemented with 10 ng/ml fibroblast growth factor 2, with and without the SB505124 inhibitor. Cells were cultured for 14 days after which gene analysis was performed. Gene expression levels were normalized to expression levels of housekeeping gene, GAPDH (shown in dotted line). Data points show mean ± SE. Statistical significance was assessed using a two-way ANOVA with Bonferroni  post-test . ** p

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Chondroinduction of Mesenchymal Stem Cells on Cellulose-Silk Composite Nanofibrous Substrates: The Role of Substrate Elasticity

    doi: 10.3389/fbioe.2020.00197

    Figure Lengend Snippet: TGFβ-driven Smad2/3 phosphorylation and stem cell chondrogenesis. hMSCs were cultured on (A) Tissue cultured plastic in chondrogenic media supplemented with transforming growth factor-β (TGFβ) and (B) composite cellulose:silk 75:25 substrates in stem cell expansion media supplemented with 10 ng/ml fibroblast growth factor 2, with and without the SB505124 inhibitor. Cells were cultured for 14 days after which gene analysis was performed. Gene expression levels were normalized to expression levels of housekeeping gene, GAPDH (shown in dotted line). Data points show mean ± SE. Statistical significance was assessed using a two-way ANOVA with Bonferroni post-test . ** p

    Article Snippet: Media was supplemented with 5 or 10 ng/ml fibroblast growth factor 2 (FGF- 2, PeproTech).

    Techniques: Cell Culture, Expressing

    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

    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