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  • 99
    Thermo Fisher dmem ko
    Dmem Ko, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 91 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore 2sb 3ct
    2sb 3ct, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Dojindo Labs fura2 am
    Fura2 Am, supplied by Dojindo Labs, used in various techniques. Bioz Stars score: 92/100, based on 66 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore fibronectin
    Fibronectin, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 20291 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Tocris sb431542
    <t>SB431542</t> exposure during early differentiation enhanced the hiPSC differentiation to corneal epithelial progenitors. Schematic outline for optimization experiment using SB431542 (A) . Quantitative real‐time polymerase chain reaction analysis of the putative limbal stem cells ( ΔNp63 ) gene for SB‐Ad3 cells from G5 subgroups on day 20 of optimization experiment following different durations of SB431542 exposure (B) . Colony forming efficiency assays for the SB431542 exposed and unexposed G5 subgroups on day 20 (C) . Data presented as mean ± SEM, n = 3. *, statistically different compared with untreated group. **, p
    Sb431542, supplied by Tocris, used in various techniques. Bioz Stars score: 99/100, based on 1892 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher 1x neaa
    <t>SB431542</t> exposure during early differentiation enhanced the hiPSC differentiation to corneal epithelial progenitors. Schematic outline for optimization experiment using SB431542 (A) . Quantitative real‐time polymerase chain reaction analysis of the putative limbal stem cells ( ΔNp63 ) gene for SB‐Ad3 cells from G5 subgroups on day 20 of optimization experiment following different durations of SB431542 exposure (B) . Colony forming efficiency assays for the SB431542 exposed and unexposed G5 subgroups on day 20 (C) . Data presented as mean ± SEM, n = 3. *, statistically different compared with untreated group. **, p
    1x Neaa, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 149 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad c1000 touch thermal cycler
    <t>SB431542</t> exposure during early differentiation enhanced the hiPSC differentiation to corneal epithelial progenitors. Schematic outline for optimization experiment using SB431542 (A) . Quantitative real‐time polymerase chain reaction analysis of the putative limbal stem cells ( ΔNp63 ) gene for SB‐Ad3 cells from G5 subgroups on day 20 of optimization experiment following different durations of SB431542 exposure (B) . Colony forming efficiency assays for the SB431542 exposed and unexposed G5 subgroups on day 20 (C) . Data presented as mean ± SEM, n = 3. *, statistically different compared with untreated group. **, p
    C1000 Touch Thermal Cycler, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 5926 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Ipsen Group syntaxin sx
    Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), <t>syntaxin</t> (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain
    Syntaxin Sx, supplied by Ipsen Group, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher ksr xf
    Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), <t>syntaxin</t> (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain
    Ksr Xf, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore a protinin
    Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), <t>syntaxin</t> (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain
    A Protinin, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Thermo Fisher atg deficient frt hygro fusion gene
    Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), <t>syntaxin</t> (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain
    Atg Deficient Frt Hygro Fusion Gene, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore β mercaptoethanol
    Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), <t>syntaxin</t> (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain
    β Mercaptoethanol, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 28107 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher β mercaptoethanol
    Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), <t>syntaxin</t> (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain
    β Mercaptoethanol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 18648 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher ion torrent proton sequencer
    Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), <t>syntaxin</t> (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain
    Ion Torrent Proton Sequencer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 423 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Millipore jak inhibitor i
    Oct4 mediates Nanog reactivation and pluripotency in somitogenesis-stage embryo explants. ( A ) Schematic diagram showing regions dissected and assays performed. ( B ) Scoring of Nanog-positive colonies from explanted TgOct4/+;rtTA/+ forebrain regions of the indicated stage embryos cultured in the presence or absence of doxycycline. Successful (+) or unsuccessful (–) EpiSC line derivation at each stage is indicated. Asterisk indicates only one Nanog-positive colony was observed after 24 hours. N/D, not determined. Number of colonies scored: E8.5+Dox, n =68; E8.5-Dox, n =53; E9.5, n =648; E10.5, n =675; E11.5, n =143; E13.5, n =220; E14.5-48 hours, n =283; E14.5-120 hours, n =428; E15.5-48 hours, n =145; E15.5-120 hours, n =660. ( C ) Nanog and Oct4 or Sox2 immunofluorescence in E8.5 and E9.5 forebrain explants. ( D ) Established EpiSC lines. Top: bright-field images showing morphology of representative lines. Bottom: Oct4 and Sox2 immunofluorescence. Box indicates the region magnified below. ( E ) mRNA expression in established EpiSC lines derived from the indicated embryonic stages. Data are mean±s.e.m. ( F ) Teratocarcinoma formation by an established EpiSC line derived from E10.5 forebrain and cultured for 1 month in <t>Jak</t> inhibitor I, and by freshly dissected forebrain explants transplanted to untreated and doxycycline-treated mice. Grafts were scored for germ layer differentiation. The presence of endodermal tissues in one of the latter tumours may result from inclusion of anterior foregut in the dissected tissue.
    Jak Inhibitor I, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 488 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore pmsf
    Oct4 mediates Nanog reactivation and pluripotency in somitogenesis-stage embryo explants. ( A ) Schematic diagram showing regions dissected and assays performed. ( B ) Scoring of Nanog-positive colonies from explanted TgOct4/+;rtTA/+ forebrain regions of the indicated stage embryos cultured in the presence or absence of doxycycline. Successful (+) or unsuccessful (–) EpiSC line derivation at each stage is indicated. Asterisk indicates only one Nanog-positive colony was observed after 24 hours. N/D, not determined. Number of colonies scored: E8.5+Dox, n =68; E8.5-Dox, n =53; E9.5, n =648; E10.5, n =675; E11.5, n =143; E13.5, n =220; E14.5-48 hours, n =283; E14.5-120 hours, n =428; E15.5-48 hours, n =145; E15.5-120 hours, n =660. ( C ) Nanog and Oct4 or Sox2 immunofluorescence in E8.5 and E9.5 forebrain explants. ( D ) Established EpiSC lines. Top: bright-field images showing morphology of representative lines. Bottom: Oct4 and Sox2 immunofluorescence. Box indicates the region magnified below. ( E ) mRNA expression in established EpiSC lines derived from the indicated embryonic stages. Data are mean±s.e.m. ( F ) Teratocarcinoma formation by an established EpiSC line derived from E10.5 forebrain and cultured for 1 month in <t>Jak</t> inhibitor I, and by freshly dissected forebrain explants transplanted to untreated and doxycycline-treated mice. Grafts were scored for germ layer differentiation. The presence of endodermal tissues in one of the latter tumours may result from inclusion of anterior foregut in the dissected tissue.
    Pmsf, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 32146 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore pepstatin a
    Oct4 mediates Nanog reactivation and pluripotency in somitogenesis-stage embryo explants. ( A ) Schematic diagram showing regions dissected and assays performed. ( B ) Scoring of Nanog-positive colonies from explanted TgOct4/+;rtTA/+ forebrain regions of the indicated stage embryos cultured in the presence or absence of doxycycline. Successful (+) or unsuccessful (–) EpiSC line derivation at each stage is indicated. Asterisk indicates only one Nanog-positive colony was observed after 24 hours. N/D, not determined. Number of colonies scored: E8.5+Dox, n =68; E8.5-Dox, n =53; E9.5, n =648; E10.5, n =675; E11.5, n =143; E13.5, n =220; E14.5-48 hours, n =283; E14.5-120 hours, n =428; E15.5-48 hours, n =145; E15.5-120 hours, n =660. ( C ) Nanog and Oct4 or Sox2 immunofluorescence in E8.5 and E9.5 forebrain explants. ( D ) Established EpiSC lines. Top: bright-field images showing morphology of representative lines. Bottom: Oct4 and Sox2 immunofluorescence. Box indicates the region magnified below. ( E ) mRNA expression in established EpiSC lines derived from the indicated embryonic stages. Data are mean±s.e.m. ( F ) Teratocarcinoma formation by an established EpiSC line derived from E10.5 forebrain and cultured for 1 month in <t>Jak</t> inhibitor I, and by freshly dissected forebrain explants transplanted to untreated and doxycycline-treated mice. Grafts were scored for germ layer differentiation. The presence of endodermal tissues in one of the latter tumours may result from inclusion of anterior foregut in the dissected tissue.
    Pepstatin A, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 7108 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore leupeptin
    Oct4 mediates Nanog reactivation and pluripotency in somitogenesis-stage embryo explants. ( A ) Schematic diagram showing regions dissected and assays performed. ( B ) Scoring of Nanog-positive colonies from explanted TgOct4/+;rtTA/+ forebrain regions of the indicated stage embryos cultured in the presence or absence of doxycycline. Successful (+) or unsuccessful (–) EpiSC line derivation at each stage is indicated. Asterisk indicates only one Nanog-positive colony was observed after 24 hours. N/D, not determined. Number of colonies scored: E8.5+Dox, n =68; E8.5-Dox, n =53; E9.5, n =648; E10.5, n =675; E11.5, n =143; E13.5, n =220; E14.5-48 hours, n =283; E14.5-120 hours, n =428; E15.5-48 hours, n =145; E15.5-120 hours, n =660. ( C ) Nanog and Oct4 or Sox2 immunofluorescence in E8.5 and E9.5 forebrain explants. ( D ) Established EpiSC lines. Top: bright-field images showing morphology of representative lines. Bottom: Oct4 and Sox2 immunofluorescence. Box indicates the region magnified below. ( E ) mRNA expression in established EpiSC lines derived from the indicated embryonic stages. Data are mean±s.e.m. ( F ) Teratocarcinoma formation by an established EpiSC line derived from E10.5 forebrain and cultured for 1 month in <t>Jak</t> inhibitor I, and by freshly dissected forebrain explants transplanted to untreated and doxycycline-treated mice. Grafts were scored for germ layer differentiation. The presence of endodermal tissues in one of the latter tumours may result from inclusion of anterior foregut in the dissected tissue.
    Leupeptin, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 18436 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Exelixis cher273 tm6 tb
    Oct4 mediates Nanog reactivation and pluripotency in somitogenesis-stage embryo explants. ( A ) Schematic diagram showing regions dissected and assays performed. ( B ) Scoring of Nanog-positive colonies from explanted TgOct4/+;rtTA/+ forebrain regions of the indicated stage embryos cultured in the presence or absence of doxycycline. Successful (+) or unsuccessful (–) EpiSC line derivation at each stage is indicated. Asterisk indicates only one Nanog-positive colony was observed after 24 hours. N/D, not determined. Number of colonies scored: E8.5+Dox, n =68; E8.5-Dox, n =53; E9.5, n =648; E10.5, n =675; E11.5, n =143; E13.5, n =220; E14.5-48 hours, n =283; E14.5-120 hours, n =428; E15.5-48 hours, n =145; E15.5-120 hours, n =660. ( C ) Nanog and Oct4 or Sox2 immunofluorescence in E8.5 and E9.5 forebrain explants. ( D ) Established EpiSC lines. Top: bright-field images showing morphology of representative lines. Bottom: Oct4 and Sox2 immunofluorescence. Box indicates the region magnified below. ( E ) mRNA expression in established EpiSC lines derived from the indicated embryonic stages. Data are mean±s.e.m. ( F ) Teratocarcinoma formation by an established EpiSC line derived from E10.5 forebrain and cultured for 1 month in <t>Jak</t> inhibitor I, and by freshly dissected forebrain explants transplanted to untreated and doxycycline-treated mice. Grafts were scored for germ layer differentiation. The presence of endodermal tissues in one of the latter tumours may result from inclusion of anterior foregut in the dissected tissue.
    Cher273 Tm6 Tb, supplied by Exelixis, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore brdu
    Oct4 mediates Nanog reactivation and pluripotency in somitogenesis-stage embryo explants. ( A ) Schematic diagram showing regions dissected and assays performed. ( B ) Scoring of Nanog-positive colonies from explanted TgOct4/+;rtTA/+ forebrain regions of the indicated stage embryos cultured in the presence or absence of doxycycline. Successful (+) or unsuccessful (–) EpiSC line derivation at each stage is indicated. Asterisk indicates only one Nanog-positive colony was observed after 24 hours. N/D, not determined. Number of colonies scored: E8.5+Dox, n =68; E8.5-Dox, n =53; E9.5, n =648; E10.5, n =675; E11.5, n =143; E13.5, n =220; E14.5-48 hours, n =283; E14.5-120 hours, n =428; E15.5-48 hours, n =145; E15.5-120 hours, n =660. ( C ) Nanog and Oct4 or Sox2 immunofluorescence in E8.5 and E9.5 forebrain explants. ( D ) Established EpiSC lines. Top: bright-field images showing morphology of representative lines. Bottom: Oct4 and Sox2 immunofluorescence. Box indicates the region magnified below. ( E ) mRNA expression in established EpiSC lines derived from the indicated embryonic stages. Data are mean±s.e.m. ( F ) Teratocarcinoma formation by an established EpiSC line derived from E10.5 forebrain and cultured for 1 month in <t>Jak</t> inhibitor I, and by freshly dissected forebrain explants transplanted to untreated and doxycycline-treated mice. Grafts were scored for germ layer differentiation. The presence of endodermal tissues in one of the latter tumours may result from inclusion of anterior foregut in the dissected tissue.
    Brdu, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 28756 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher fibronectin
    CaSm induction promotes migration and invasion and alters metastatic gene expression. Tet-on driver and tet-on CaSm Panc-1 cells, previously grown in 1 μg/ml doxycycline for 4–8 weeks, were plated in serum-free DMEM onto the top chamber of <t>fibronectin-coated</t> transwell membranes ( a ) or rehydrated Matrigel transwell chambers ( b ). DMEM supplemented with 10% FBS was used as the chemoattractant in the bottom wells. The chambers were incubated at 37 °C for 4 h ( a ) and 48 h ( b ) before transwell cells were stained and quantified by counting random fields at × 40 magnification. Represented is mean and s.e.m. ( c ) Real-time PCR was performed to validate the microarray with samples normalized to GAPDH as a loading control. * P
    Fibronectin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4592 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Selleck Chemicals forskolin
    TGF-β1/ALK5 signaling induces Prg4 expression via the PKA-CREB signaling pathway in chondrocytes A, B, G and H. Femoral head cartilage isolated from Prg4 GFPCreERT2/+ mice was pretreated with selective inhibitor for 30 min, followed by an additional 24 h incubation with TGF-β1, inhibitor, or a combination of both. qRT-PCR and western blotting were performed to detect the expression of Prg4 mRNA (A and G, two-way ANOVA followed by Tukey’s test, n = 3 femoral head cartilage per group) and the GFP protein expression (B and H, quantitative data were shown in the lower panel, two-way ANOVA followed by Tukey’s test, n = 3 femoral head cartilage per group), respectively. C and D, Primary chondrocytes isolated from Cre-negative mice were pretreated with 10 μM H89 (C) or 1 μM SB-505124 (D) for 30 min, followed by an additional 30 min incubation of 10 ng/ml TGF-β1,10 μM H89/1 μM SB-505124, or a combination of both. Western blotting was performed to detect the pCREB and pSmad3 protein expressions. Quantitative data were shown in the right panel (two-way ANOVA followed by Tukey’s test, n = 4 mice per group). E, Western blotting analysis of pCREB in Cre-negative or Alk5 -deficient primary chondrocytes treated with TGF-β1 for 30 min. Quantitative data were shown in the right panel (two-way ANOVA followed by Tukey’s test, n = 4 mice per group). F, IHC analysis of pCREB protein expression in articular cartilage of 2-month-old Alk5 cKO mice and Cre-negative control mice. Quantitative data were shown in the right panel (The percentage of positive cells in Cre-negative mice was defined as 1, student’s unpaired t -test, n = 4 mice per group). I, qRT-PCR analysis of Prg4 mRNA expression in Cre-negative or Alk5 -deficient femoral head cartilage treated with increasing concentrations of <t>forskolin</t> (two-way ANOVA followed by Tukey’s test, n = 3 femoral head cartilage per group). J. Schematic illustration of the mechanisms of ALK5 signaling in the maintenance of articular cartilage. Data were expressed as the mean ± 95% confidence intervals. In F, symbols represent individual mice. In A, B, C, D, E, G and H, symbols represent three technical replicates.
    Forskolin, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 95/100, based on 73 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech human recombinant egf
    Signaling pathway studies of <t>EGF</t> regulation of <t>hNHE8</t> gene expression. Caco-2 cells were transfected with promoter construct pGL3/-32 bp and pretreated with various inhibitors for 2 h before EGF was added. Relative change is shown as the ratio of luciferase activity in EGF-treated cells to luciferase activity in vehicle-treated cells in the presence or absence of various inhibitors. Results are means ± SE from 4 independent experiments. * P
    Human Recombinant Egf, supplied by PeproTech, used in various techniques. Bioz Stars score: 99/100, based on 310 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen plasmid isolation kit
    Signaling pathway studies of <t>EGF</t> regulation of <t>hNHE8</t> gene expression. Caco-2 cells were transfected with promoter construct pGL3/-32 bp and pretreated with various inhibitors for 2 h before EGF was added. Relative change is shown as the ratio of luciferase activity in EGF-treated cells to luciferase activity in vehicle-treated cells in the presence or absence of various inhibitors. Results are means ± SE from 4 independent experiments. * P
    Plasmid Isolation Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 805 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher proton sequencer supplementary
    Signaling pathway studies of <t>EGF</t> regulation of <t>hNHE8</t> gene expression. Caco-2 cells were transfected with promoter construct pGL3/-32 bp and pretreated with various inhibitors for 2 h before EGF was added. Relative change is shown as the ratio of luciferase activity in EGF-treated cells to luciferase activity in vehicle-treated cells in the presence or absence of various inhibitors. Results are means ± SE from 4 independent experiments. * P
    Proton Sequencer Supplementary, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    SB431542 exposure during early differentiation enhanced the hiPSC differentiation to corneal epithelial progenitors. Schematic outline for optimization experiment using SB431542 (A) . Quantitative real‐time polymerase chain reaction analysis of the putative limbal stem cells ( ΔNp63 ) gene for SB‐Ad3 cells from G5 subgroups on day 20 of optimization experiment following different durations of SB431542 exposure (B) . Colony forming efficiency assays for the SB431542 exposed and unexposed G5 subgroups on day 20 (C) . Data presented as mean ± SEM, n = 3. *, statistically different compared with untreated group. **, p

    Journal: Stem Cells (Dayton, Ohio)

    Article Title: Differences in the Activity of Endogenous Bone Morphogenetic Protein Signaling Impact on the Ability of Induced Pluripotent Stem Cells to Differentiate to Corneal Epithelial‐Like Cells

    doi: 10.1002/stem.2750

    Figure Lengend Snippet: SB431542 exposure during early differentiation enhanced the hiPSC differentiation to corneal epithelial progenitors. Schematic outline for optimization experiment using SB431542 (A) . Quantitative real‐time polymerase chain reaction analysis of the putative limbal stem cells ( ΔNp63 ) gene for SB‐Ad3 cells from G5 subgroups on day 20 of optimization experiment following different durations of SB431542 exposure (B) . Colony forming efficiency assays for the SB431542 exposed and unexposed G5 subgroups on day 20 (C) . Data presented as mean ± SEM, n = 3. *, statistically different compared with untreated group. **, p

    Article Snippet: SB431542 (10 µM) was added for 1, 2, and 3 days to the differentiation media in combination with BMP4, RA, and EGF as detailed in Figure A. qRT‐PCR analysis at day 20 indicated the highest expression of ΔNp63 in groups treated for 2 and 3 days with SB431542 (Fig. B).

    Techniques: Real-time Polymerase Chain Reaction

    Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), syntaxin (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain

    Journal: Biophysical Journal

    Article Title: Simulations Reveal Multiple Intermediates in the Unzipping Mechanism of Neuronal SNARE Complex

    doi: 10.1016/j.bpj.2018.08.043

    Figure Lengend Snippet: Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), syntaxin (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain

    Article Snippet: Synaptobrevin (Sb) and syntaxin (Sx) both contribute to this complex with one α -helix, whereas SNAP25 consists of two α -helical motifs connected by an unstructured linker loop.

    Techniques:

    Oct4 mediates Nanog reactivation and pluripotency in somitogenesis-stage embryo explants. ( A ) Schematic diagram showing regions dissected and assays performed. ( B ) Scoring of Nanog-positive colonies from explanted TgOct4/+;rtTA/+ forebrain regions of the indicated stage embryos cultured in the presence or absence of doxycycline. Successful (+) or unsuccessful (–) EpiSC line derivation at each stage is indicated. Asterisk indicates only one Nanog-positive colony was observed after 24 hours. N/D, not determined. Number of colonies scored: E8.5+Dox, n =68; E8.5-Dox, n =53; E9.5, n =648; E10.5, n =675; E11.5, n =143; E13.5, n =220; E14.5-48 hours, n =283; E14.5-120 hours, n =428; E15.5-48 hours, n =145; E15.5-120 hours, n =660. ( C ) Nanog and Oct4 or Sox2 immunofluorescence in E8.5 and E9.5 forebrain explants. ( D ) Established EpiSC lines. Top: bright-field images showing morphology of representative lines. Bottom: Oct4 and Sox2 immunofluorescence. Box indicates the region magnified below. ( E ) mRNA expression in established EpiSC lines derived from the indicated embryonic stages. Data are mean±s.e.m. ( F ) Teratocarcinoma formation by an established EpiSC line derived from E10.5 forebrain and cultured for 1 month in Jak inhibitor I, and by freshly dissected forebrain explants transplanted to untreated and doxycycline-treated mice. Grafts were scored for germ layer differentiation. The presence of endodermal tissues in one of the latter tumours may result from inclusion of anterior foregut in the dissected tissue.

    Journal: Development (Cambridge, England)

    Article Title: The developmental dismantling of pluripotency is reversed by ectopic Oct4 expression

    doi: 10.1242/dev.078071

    Figure Lengend Snippet: Oct4 mediates Nanog reactivation and pluripotency in somitogenesis-stage embryo explants. ( A ) Schematic diagram showing regions dissected and assays performed. ( B ) Scoring of Nanog-positive colonies from explanted TgOct4/+;rtTA/+ forebrain regions of the indicated stage embryos cultured in the presence or absence of doxycycline. Successful (+) or unsuccessful (–) EpiSC line derivation at each stage is indicated. Asterisk indicates only one Nanog-positive colony was observed after 24 hours. N/D, not determined. Number of colonies scored: E8.5+Dox, n =68; E8.5-Dox, n =53; E9.5, n =648; E10.5, n =675; E11.5, n =143; E13.5, n =220; E14.5-48 hours, n =283; E14.5-120 hours, n =428; E15.5-48 hours, n =145; E15.5-120 hours, n =660. ( C ) Nanog and Oct4 or Sox2 immunofluorescence in E8.5 and E9.5 forebrain explants. ( D ) Established EpiSC lines. Top: bright-field images showing morphology of representative lines. Bottom: Oct4 and Sox2 immunofluorescence. Box indicates the region magnified below. ( E ) mRNA expression in established EpiSC lines derived from the indicated embryonic stages. Data are mean±s.e.m. ( F ) Teratocarcinoma formation by an established EpiSC line derived from E10.5 forebrain and cultured for 1 month in Jak inhibitor I, and by freshly dissected forebrain explants transplanted to untreated and doxycycline-treated mice. Grafts were scored for germ layer differentiation. The presence of endodermal tissues in one of the latter tumours may result from inclusion of anterior foregut in the dissected tissue.

    Article Snippet: When added, chemical inhibitors were used at these concentrations: PD0325901 (Stemgent), 1 μM; SB 431542 (Sigma), 10 μM; JAK inhibitor I (Calbiochem 420099), 0.6 mM.

    Techniques: Cell Culture, Immunofluorescence, Expressing, Derivative Assay, Mouse Assay

    CaSm induction promotes migration and invasion and alters metastatic gene expression. Tet-on driver and tet-on CaSm Panc-1 cells, previously grown in 1 μg/ml doxycycline for 4–8 weeks, were plated in serum-free DMEM onto the top chamber of fibronectin-coated transwell membranes ( a ) or rehydrated Matrigel transwell chambers ( b ). DMEM supplemented with 10% FBS was used as the chemoattractant in the bottom wells. The chambers were incubated at 37 °C for 4 h ( a ) and 48 h ( b ) before transwell cells were stained and quantified by counting random fields at × 40 magnification. Represented is mean and s.e.m. ( c ) Real-time PCR was performed to validate the microarray with samples normalized to GAPDH as a loading control. * P

    Journal: Oncogenesis

    Article Title: The CaSm (LSm1) oncogene promotes transformation, chemoresistance and metastasis of pancreatic cancer cells

    doi: 10.1038/oncsis.2015.45

    Figure Lengend Snippet: CaSm induction promotes migration and invasion and alters metastatic gene expression. Tet-on driver and tet-on CaSm Panc-1 cells, previously grown in 1 μg/ml doxycycline for 4–8 weeks, were plated in serum-free DMEM onto the top chamber of fibronectin-coated transwell membranes ( a ) or rehydrated Matrigel transwell chambers ( b ). DMEM supplemented with 10% FBS was used as the chemoattractant in the bottom wells. The chambers were incubated at 37 °C for 4 h ( a ) and 48 h ( b ) before transwell cells were stained and quantified by counting random fields at × 40 magnification. Represented is mean and s.e.m. ( c ) Real-time PCR was performed to validate the microarray with samples normalized to GAPDH as a loading control. * P

    Article Snippet: Migration assay Twenty-four-well cell culture inserts with 8.0 um pores (BD Biosciences) were coated with 15 μg/ml fibronectin (ThermoScientific) overnight at 4 °C.

    Techniques: Migration, Expressing, Incubation, Staining, Real-time Polymerase Chain Reaction, Microarray

    TGF-β1/ALK5 signaling induces Prg4 expression via the PKA-CREB signaling pathway in chondrocytes A, B, G and H. Femoral head cartilage isolated from Prg4 GFPCreERT2/+ mice was pretreated with selective inhibitor for 30 min, followed by an additional 24 h incubation with TGF-β1, inhibitor, or a combination of both. qRT-PCR and western blotting were performed to detect the expression of Prg4 mRNA (A and G, two-way ANOVA followed by Tukey’s test, n = 3 femoral head cartilage per group) and the GFP protein expression (B and H, quantitative data were shown in the lower panel, two-way ANOVA followed by Tukey’s test, n = 3 femoral head cartilage per group), respectively. C and D, Primary chondrocytes isolated from Cre-negative mice were pretreated with 10 μM H89 (C) or 1 μM SB-505124 (D) for 30 min, followed by an additional 30 min incubation of 10 ng/ml TGF-β1,10 μM H89/1 μM SB-505124, or a combination of both. Western blotting was performed to detect the pCREB and pSmad3 protein expressions. Quantitative data were shown in the right panel (two-way ANOVA followed by Tukey’s test, n = 4 mice per group). E, Western blotting analysis of pCREB in Cre-negative or Alk5 -deficient primary chondrocytes treated with TGF-β1 for 30 min. Quantitative data were shown in the right panel (two-way ANOVA followed by Tukey’s test, n = 4 mice per group). F, IHC analysis of pCREB protein expression in articular cartilage of 2-month-old Alk5 cKO mice and Cre-negative control mice. Quantitative data were shown in the right panel (The percentage of positive cells in Cre-negative mice was defined as 1, student’s unpaired t -test, n = 4 mice per group). I, qRT-PCR analysis of Prg4 mRNA expression in Cre-negative or Alk5 -deficient femoral head cartilage treated with increasing concentrations of forskolin (two-way ANOVA followed by Tukey’s test, n = 3 femoral head cartilage per group). J. Schematic illustration of the mechanisms of ALK5 signaling in the maintenance of articular cartilage. Data were expressed as the mean ± 95% confidence intervals. In F, symbols represent individual mice. In A, B, C, D, E, G and H, symbols represent three technical replicates.

    Journal: Osteoarthritis and cartilage

    Article Title: Cartilage-specific deletion of Alk5 gene results in a progressive osteoarthritis-like phenotype in mice

    doi: 10.1016/j.joca.2017.07.010

    Figure Lengend Snippet: TGF-β1/ALK5 signaling induces Prg4 expression via the PKA-CREB signaling pathway in chondrocytes A, B, G and H. Femoral head cartilage isolated from Prg4 GFPCreERT2/+ mice was pretreated with selective inhibitor for 30 min, followed by an additional 24 h incubation with TGF-β1, inhibitor, or a combination of both. qRT-PCR and western blotting were performed to detect the expression of Prg4 mRNA (A and G, two-way ANOVA followed by Tukey’s test, n = 3 femoral head cartilage per group) and the GFP protein expression (B and H, quantitative data were shown in the lower panel, two-way ANOVA followed by Tukey’s test, n = 3 femoral head cartilage per group), respectively. C and D, Primary chondrocytes isolated from Cre-negative mice were pretreated with 10 μM H89 (C) or 1 μM SB-505124 (D) for 30 min, followed by an additional 30 min incubation of 10 ng/ml TGF-β1,10 μM H89/1 μM SB-505124, or a combination of both. Western blotting was performed to detect the pCREB and pSmad3 protein expressions. Quantitative data were shown in the right panel (two-way ANOVA followed by Tukey’s test, n = 4 mice per group). E, Western blotting analysis of pCREB in Cre-negative or Alk5 -deficient primary chondrocytes treated with TGF-β1 for 30 min. Quantitative data were shown in the right panel (two-way ANOVA followed by Tukey’s test, n = 4 mice per group). F, IHC analysis of pCREB protein expression in articular cartilage of 2-month-old Alk5 cKO mice and Cre-negative control mice. Quantitative data were shown in the right panel (The percentage of positive cells in Cre-negative mice was defined as 1, student’s unpaired t -test, n = 4 mice per group). I, qRT-PCR analysis of Prg4 mRNA expression in Cre-negative or Alk5 -deficient femoral head cartilage treated with increasing concentrations of forskolin (two-way ANOVA followed by Tukey’s test, n = 3 femoral head cartilage per group). J. Schematic illustration of the mechanisms of ALK5 signaling in the maintenance of articular cartilage. Data were expressed as the mean ± 95% confidence intervals. In F, symbols represent individual mice. In A, B, C, D, E, G and H, symbols represent three technical replicates.

    Article Snippet: Femoral head cartilage isolated from Alk5Col2ERT2 and Cre-negative mice was cultured in tissue culture method and treated with 1 μM 4-hydroxy tamoxifen (4-OH TM) (Sigma–Aldrich) for 72 h to drive Cre-mediated recombination and delete Alk5 , then treated with TGF-β1 (Peprotech) or forskolin (Selleck) for indicated time.

    Techniques: Expressing, Isolation, Mouse Assay, Incubation, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Negative Control

    Signaling pathway studies of EGF regulation of hNHE8 gene expression. Caco-2 cells were transfected with promoter construct pGL3/-32 bp and pretreated with various inhibitors for 2 h before EGF was added. Relative change is shown as the ratio of luciferase activity in EGF-treated cells to luciferase activity in vehicle-treated cells in the presence or absence of various inhibitors. Results are means ± SE from 4 independent experiments. * P

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Epidermal growth factor inhibits intestinal NHE8 expression via reducing its basal transcription

    doi: 10.1152/ajpcell.00081.2010

    Figure Lengend Snippet: Signaling pathway studies of EGF regulation of hNHE8 gene expression. Caco-2 cells were transfected with promoter construct pGL3/-32 bp and pretreated with various inhibitors for 2 h before EGF was added. Relative change is shown as the ratio of luciferase activity in EGF-treated cells to luciferase activity in vehicle-treated cells in the presence or absence of various inhibitors. Results are means ± SE from 4 independent experiments. * P

    Article Snippet: To test the effect of EGF on hNHE8 promoter activity, transfected cells were treated with 100 ng/ml human recombinant EGF for 18 h before promoter reporter assay.

    Techniques: Expressing, Transfection, Construct, Luciferase, Activity Assay