basic fibroblast growth factor bfgf 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 1068 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
    Bfgf, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2455 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore fibroblast growth factor 2
    Neurogenic ability of non-pigmented CE and Müller stem cells cultured under differentiating conditions for 7 days (A) (i) Morphology of CE and Müller cells cultured under normal culture conditions in the presence of 10% serum. Non-pigmented CE cells were maintained with EGF. (ii) Under adherent conditions in the presence of ECM gel and FGF2, non-pigmented CE cells maintained their epithelial cell morphology, however, a few flattened cells were visible. In contrast, Müller stem cells acquired a neural-like morphology (white arrows). (iii) In the presence of DAPT and FGF2, CE cells ceased to proliferate and did not adopt a neural morphology. However under these conditions a greater proportion of Müller stem cells adopted a neural-like morphology. (Scale bar represents 100 μm.) (B) Western blotting of cell lysates revealed that CE cell preparations cultured under differentiating conditions (CE 6165, CE 6213 and CE 6334) did not express BRN3B, a transcription factor present in post-mitotic retinal ganglion cell precursors. However, increased expression of this protein was observed in Müller stem cells (MIO-M2, MIO-M5 and MIO-M7) cultured under similar conditions. (C) Examination of mRNA for photoreceptor gene expression revealed that in the presence of FGF2 non-pigmented CE cells and Müller stem cells expressed  NRL  and Rhodopsin, but that only Müller cells expressed S-Opsin. (CE = Ciliary Epithelium, FGF2 = Fibroblast Growth Factor 2.)
    Fibroblast Growth Factor 2, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 515 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 882 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher basic fibroblast growth factor bfgf
    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; <t>bFGF,</t> basic fibroblast growth factor; <t>EGF,</t> epidermal growth factor.
    Basic Fibroblast Growth Factor Bfgf, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 964 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher fibroblast growth factor
    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; <t>bFGF,</t> basic fibroblast growth factor; <t>EGF,</t> epidermal growth factor.
    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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    PeproTech basic fibroblast growth factor
    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; <t>bFGF,</t> basic fibroblast growth factor; <t>EGF,</t> epidermal growth factor.
    Basic Fibroblast Growth Factor, supplied by PeproTech, used in various techniques. Bioz Stars score: 99/100, based on 3182 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    FUJIFILM basic fibroblast growth factor
    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; <t>bFGF,</t> basic fibroblast growth factor; <t>EGF,</t> epidermal growth factor.
    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 fgf basic aa 1 155 recombinant human protein
    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; <t>bFGF,</t> basic fibroblast growth factor; <t>EGF,</t> epidermal growth factor.
    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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    FUJIFILM recombinant human basic fibroblast growth factor
    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; <t>bFGF,</t> basic fibroblast growth factor; <t>EGF,</t> epidermal growth factor.
    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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    98
    Thermo Fisher fgf basic aa 10 155 recombinant human protein
    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; <t>bFGF,</t> basic fibroblast growth factor; <t>EGF,</t> epidermal growth factor.
    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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    Thermo Fisher human basic fibroblast growth factor
    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; <t>bFGF,</t> basic fibroblast growth factor; <t>EGF,</t> epidermal growth factor.
    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
    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; <t>bFGF,</t> basic fibroblast growth factor; <t>EGF,</t> epidermal growth factor.
    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
    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; <t>bFGF,</t> basic fibroblast growth factor; <t>EGF,</t> epidermal growth factor.
    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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    93
    Thermo Fisher bfgf
    <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.
    Bfgf, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 3851 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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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

    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

    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

    Neurogenic ability of non-pigmented CE and Müller stem cells cultured under differentiating conditions for 7 days (A) (i) Morphology of CE and Müller cells cultured under normal culture conditions in the presence of 10% serum. Non-pigmented CE cells were maintained with EGF. (ii) Under adherent conditions in the presence of ECM gel and FGF2, non-pigmented CE cells maintained their epithelial cell morphology, however, a few flattened cells were visible. In contrast, Müller stem cells acquired a neural-like morphology (white arrows). (iii) In the presence of DAPT and FGF2, CE cells ceased to proliferate and did not adopt a neural morphology. However under these conditions a greater proportion of Müller stem cells adopted a neural-like morphology. (Scale bar represents 100 μm.) (B) Western blotting of cell lysates revealed that CE cell preparations cultured under differentiating conditions (CE 6165, CE 6213 and CE 6334) did not express BRN3B, a transcription factor present in post-mitotic retinal ganglion cell precursors. However, increased expression of this protein was observed in Müller stem cells (MIO-M2, MIO-M5 and MIO-M7) cultured under similar conditions. (C) Examination of mRNA for photoreceptor gene expression revealed that in the presence of FGF2 non-pigmented CE cells and Müller stem cells expressed  NRL  and Rhodopsin, but that only Müller cells expressed S-Opsin. (CE = Ciliary Epithelium, FGF2 = Fibroblast Growth Factor 2.)

    Journal: Experimental Eye Research

    Article Title: Differences between the neurogenic and proliferative abilities of M?ller glia with stem cell characteristics and the ciliary epithelium from the adult human eye

    doi: 10.1016/j.exer.2011.09.015

    Figure Lengend Snippet: Neurogenic ability of non-pigmented CE and Müller stem cells cultured under differentiating conditions for 7 days (A) (i) Morphology of CE and Müller cells cultured under normal culture conditions in the presence of 10% serum. Non-pigmented CE cells were maintained with EGF. (ii) Under adherent conditions in the presence of ECM gel and FGF2, non-pigmented CE cells maintained their epithelial cell morphology, however, a few flattened cells were visible. In contrast, Müller stem cells acquired a neural-like morphology (white arrows). (iii) In the presence of DAPT and FGF2, CE cells ceased to proliferate and did not adopt a neural morphology. However under these conditions a greater proportion of Müller stem cells adopted a neural-like morphology. (Scale bar represents 100 μm.) (B) Western blotting of cell lysates revealed that CE cell preparations cultured under differentiating conditions (CE 6165, CE 6213 and CE 6334) did not express BRN3B, a transcription factor present in post-mitotic retinal ganglion cell precursors. However, increased expression of this protein was observed in Müller stem cells (MIO-M2, MIO-M5 and MIO-M7) cultured under similar conditions. (C) Examination of mRNA for photoreceptor gene expression revealed that in the presence of FGF2 non-pigmented CE cells and Müller stem cells expressed NRL and Rhodopsin, but that only Müller cells expressed S-Opsin. (CE = Ciliary Epithelium, FGF2 = Fibroblast Growth Factor 2.)

    Article Snippet: Cells were cultured in DMEM medium containing 5% FCS, fibroblast growth factor 2 (FGF2/bFGF-40 ng/ml, Sigma) and DAPT (N-[N-(3,5-Difluorophenacetyl)-l -alanyl]-S-phenylglycine t-butyl ester, Sigma) diluted in DMSO to a concentration of 50 μM/ml.

    Techniques: Cell Culture, Western Blot, 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

    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

    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

    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