gene exp vegfa hs00173626 m1  (Thermo Fisher)


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    Structured Review

    Thermo Fisher gene exp vegfa hs00173626 m1
    In vitro tube formation by hAMSC requires Notch1 expression . Phase contrast photomicrographs ( a ) of untransduced (left); αNotch1 shRNA transduced (middle) and mock ΦshRNA transduced (right) PECAM:PL-G/RL-R-tTK-hAMSCs incubated in Matrigel for 4 hours. ( b ) RNA extracted from cells in matrigel was assayed by RT-PCR to determine transcription the levels of the indicated endothelial (ILK, SDF1, EGR3,PECAM-1, <t>VEGFA-1,</t> Notch-1) and bone BGLAP/Octeocalcin differentiation markers. The histogram shows % fold change relative to the control un-transduced PECAM:PL-G/RL-R-tTK-hAMSCs. * P
    Gene Exp Vegfa Hs00173626 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    gene exp vegfa hs00173626 m1 - by Bioz Stars, 2021-03
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    Images

    1) Product Images from "Endothelial Differentiation of Adipose Tissue-Derived Mesenchymal Stromal Cells in Glioma Tumors: Implications for Cell-Based Therapy"

    Article Title: Endothelial Differentiation of Adipose Tissue-Derived Mesenchymal Stromal Cells in Glioma Tumors: Implications for Cell-Based Therapy

    Journal: Molecular Therapy

    doi: 10.1038/mt.2013.145

    In vitro tube formation by hAMSC requires Notch1 expression . Phase contrast photomicrographs ( a ) of untransduced (left); αNotch1 shRNA transduced (middle) and mock ΦshRNA transduced (right) PECAM:PL-G/RL-R-tTK-hAMSCs incubated in Matrigel for 4 hours. ( b ) RNA extracted from cells in matrigel was assayed by RT-PCR to determine transcription the levels of the indicated endothelial (ILK, SDF1, EGR3,PECAM-1, VEGFA-1, Notch-1) and bone BGLAP/Octeocalcin differentiation markers. The histogram shows % fold change relative to the control un-transduced PECAM:PL-G/RL-R-tTK-hAMSCs. * P
    Figure Legend Snippet: In vitro tube formation by hAMSC requires Notch1 expression . Phase contrast photomicrographs ( a ) of untransduced (left); αNotch1 shRNA transduced (middle) and mock ΦshRNA transduced (right) PECAM:PL-G/RL-R-tTK-hAMSCs incubated in Matrigel for 4 hours. ( b ) RNA extracted from cells in matrigel was assayed by RT-PCR to determine transcription the levels of the indicated endothelial (ILK, SDF1, EGR3,PECAM-1, VEGFA-1, Notch-1) and bone BGLAP/Octeocalcin differentiation markers. The histogram shows % fold change relative to the control un-transduced PECAM:PL-G/RL-R-tTK-hAMSCs. * P

    Techniques Used: In Vitro, Expressing, shRNA, Incubation, Reverse Transcription Polymerase Chain Reaction

    2) Product Images from "Novel TPO receptor agonist TA-316 contributes to platelet biogenesis from human iPS cells"

    Article Title: Novel TPO receptor agonist TA-316 contributes to platelet biogenesis from human iPS cells

    Journal: Blood Advances

    doi: 10.1182/bloodadvances.2016000844

    An inhibitor of FGF or VEGF receptors eliminates the superior effect of TA-316 on imMKCL proliferation. imMKCLs cultured with rhTPO (50 ng/mL), TA-316 (200 ng/mL), eltrombopag (1000 ng/mL), or DMSO (0.05%). (A) Cells were collected on day 11 (genes on). mRNA expressions of VEGFA and FGF2 (genes on) were evaluated using Taqman PCR assays. GAPDH served as an internal control. (B) FGFR antagonist PD173074 (100 nM; WAKO), VEGFR antagonist axitinib (5 nM; Toronto Research Chemicals, Toronto, Canada), or epithelial growth factor receptor antagonist gefitinib (1 μM, Santa Cruz Biotechnology, Dallas, TX) was added to the imMKCLs, which were cultured with TA-316 (200 ng/mL) for 4 days (genes on). Cell proliferation was assessed based on 0.4% Trypan blue staining (Thermo Fisher Scientific). Data represent the mean and SD; * P
    Figure Legend Snippet: An inhibitor of FGF or VEGF receptors eliminates the superior effect of TA-316 on imMKCL proliferation. imMKCLs cultured with rhTPO (50 ng/mL), TA-316 (200 ng/mL), eltrombopag (1000 ng/mL), or DMSO (0.05%). (A) Cells were collected on day 11 (genes on). mRNA expressions of VEGFA and FGF2 (genes on) were evaluated using Taqman PCR assays. GAPDH served as an internal control. (B) FGFR antagonist PD173074 (100 nM; WAKO), VEGFR antagonist axitinib (5 nM; Toronto Research Chemicals, Toronto, Canada), or epithelial growth factor receptor antagonist gefitinib (1 μM, Santa Cruz Biotechnology, Dallas, TX) was added to the imMKCLs, which were cultured with TA-316 (200 ng/mL) for 4 days (genes on). Cell proliferation was assessed based on 0.4% Trypan blue staining (Thermo Fisher Scientific). Data represent the mean and SD; * P

    Techniques Used: Cell Culture, Polymerase Chain Reaction, Staining

    3) Product Images from "Kaposi's Sarcoma Herpesvirus K15 Protein Contributes to Virus-Induced Angiogenesis by Recruiting PLC?1 and Activating NFAT1-dependent RCAN1 Expression"

    Article Title: Kaposi's Sarcoma Herpesvirus K15 Protein Contributes to Virus-Induced Angiogenesis by Recruiting PLC?1 and Activating NFAT1-dependent RCAN1 Expression

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1002927

    K15 dependent cellular gene expression in HUVECs. (A) Microarray analysis of cellular mRNAs upregulated in HUVECs by K15 overexpression or after KSHV infection. HUVECs were transduced with a K15-expression vector (lanes 1–2) or infected with either KSHVwt or KSHVΔK15 (lanes 3–4). Depicted are the top-ranking 50 transcripts most strongly upregulated by K15 overexpression, showing an at least two-fold induction by K15 overexpression in each of two experiments performed using cells from two healthy donors (lanes 1–2). Asterisks next to fold change values within heatmap lanes 3–4 indicate genes that were induced more than two-fold in KSHVwt-infected cells compared to cells mock infected with heat-inactivated virus. Empty cells in the heatmap correspond to undetectable mRNA levels in both of the two samples compared in each lane. Arrows indicate cellular genes, inducible upon KSHVwt infection with more than two-fold elevated expression in KSHVwt- relative to KSHVΔK15-infected cells in each of the two experiments performed. (B) Quantitative PCR analysis of K15 transcript in mRNA derived from KSHVwt- or KSHVΔK15-infected cells used for the microarray experiment. (C) Quantitative PCR analysis of RCAN1/DSCR1 and VEGFA transcripts in HUVECs transduced with a retroviral K15 expression vector or with an empty control vector. (D) HUVECs from two healthy donors (Exp1–2) were infected with heat-inactivated, KSHVwt, or KSHVΔK15 and were lysed 4 days after infection. RNA was extracted, reversely transcribed and subjected to quantitative TaqMan-based PCR analysis. mRNA fold change values of KSHVwt-infected versus mock-infected (with heat-inactivated KSHVwt = HI) cells were depicted in black, values of KSHVΔK15- infected versus mock-infected cells were depicted in red. Input mRNA samples are identical to samples used in microarray experiments (compare panel A lanes 3–4).
    Figure Legend Snippet: K15 dependent cellular gene expression in HUVECs. (A) Microarray analysis of cellular mRNAs upregulated in HUVECs by K15 overexpression or after KSHV infection. HUVECs were transduced with a K15-expression vector (lanes 1–2) or infected with either KSHVwt or KSHVΔK15 (lanes 3–4). Depicted are the top-ranking 50 transcripts most strongly upregulated by K15 overexpression, showing an at least two-fold induction by K15 overexpression in each of two experiments performed using cells from two healthy donors (lanes 1–2). Asterisks next to fold change values within heatmap lanes 3–4 indicate genes that were induced more than two-fold in KSHVwt-infected cells compared to cells mock infected with heat-inactivated virus. Empty cells in the heatmap correspond to undetectable mRNA levels in both of the two samples compared in each lane. Arrows indicate cellular genes, inducible upon KSHVwt infection with more than two-fold elevated expression in KSHVwt- relative to KSHVΔK15-infected cells in each of the two experiments performed. (B) Quantitative PCR analysis of K15 transcript in mRNA derived from KSHVwt- or KSHVΔK15-infected cells used for the microarray experiment. (C) Quantitative PCR analysis of RCAN1/DSCR1 and VEGFA transcripts in HUVECs transduced with a retroviral K15 expression vector or with an empty control vector. (D) HUVECs from two healthy donors (Exp1–2) were infected with heat-inactivated, KSHVwt, or KSHVΔK15 and were lysed 4 days after infection. RNA was extracted, reversely transcribed and subjected to quantitative TaqMan-based PCR analysis. mRNA fold change values of KSHVwt-infected versus mock-infected (with heat-inactivated KSHVwt = HI) cells were depicted in black, values of KSHVΔK15- infected versus mock-infected cells were depicted in red. Input mRNA samples are identical to samples used in microarray experiments (compare panel A lanes 3–4).

    Techniques Used: Expressing, Microarray, Over Expression, Infection, Transduction, Plasmid Preparation, Real-time Polymerase Chain Reaction, Derivative Assay, Polymerase Chain Reaction

    4) Product Images from "Bioceramic-Mediated Trophic Factor Secretion by Mesenchymal Stem Cells Enhances In Vitro Endothelial Cell Persistence and In Vivo Angiogenesis"

    Article Title: Bioceramic-Mediated Trophic Factor Secretion by Mesenchymal Stem Cells Enhances In Vitro Endothelial Cell Persistence and In Vivo Angiogenesis

    Journal: Tissue Engineering. Part A

    doi: 10.1089/ten.tea.2011.0127

    Quantitative polymerase chain reaction was used to analyze proangiogenic gene expression by MSCs on various scaffolds. (A) VEGFA , (B) PDGF , (C) FGF1 , and (D) FGF2 expression were normalized to RPL13 expression and data were represented as ΔCt.
    Figure Legend Snippet: Quantitative polymerase chain reaction was used to analyze proangiogenic gene expression by MSCs on various scaffolds. (A) VEGFA , (B) PDGF , (C) FGF1 , and (D) FGF2 expression were normalized to RPL13 expression and data were represented as ΔCt.

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing

    5) Product Images from "Focal Gains of Vascular Endothelial Growth Factor A and Molecular Classification of Hepatocellular Carcinoma"

    Article Title: Focal Gains of Vascular Endothelial Growth Factor A and Molecular Classification of Hepatocellular Carcinoma

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-08-0742

    High-level gains at 6p21 lead to VEGFA overexpression (A) Significance Analysis of Microarrays identified overexpression of two genes among the 4 tumors with 6p21 amplifications. Each point represents the observed t -statistic, plotted against its expected score from permutation tests. The most significant gene is VEGFA , followed by TMEM63B (indicated in purple). The dashed lines indicate the confidence interval for the random, null distribution. (B) VEGFA or (C) TMEM63B is overexpressed in tumors with high-level gains of 6p21. Each of the 91 points represents the corresponding copy number and expression level for a single tumor. The horizontal axis indicates the median inferred copy number for 68 single nucleotide polymorphism probes in the minimal overlapping region at 6p21. The vertical axis indicates the log2 ratio between the expression level of each tumor, compared to the median of 10 normal liver samples on the Affymetrix U133 Plus 2 array. (D) Higher VEGFA expression is associated with increased copy numbers in an independent cohort. Each of the 45 points represents a single tumor chosen from the panel of 210 tumors evaluated by fluorescence in situ hybridization. The horizontal axis indicates the average number of VEGFA probes per nucleus. The vertical axis indicates the negative ΔΔ C t values of VEGFA normalized to ACTB , compared to the median of 5 uninvolved, cirrhotic tissue samples. Tumors with more than 4 copies of VEGFA are shown in red ( n = 10), tumors with gains of chromosome 6 are shown in black ( n = 19), and diploid tumors are shown in grey ( n = 16). The average negative ΔΔ C t value for each of these three classes is plotted with a dashed blue line.
    Figure Legend Snippet: High-level gains at 6p21 lead to VEGFA overexpression (A) Significance Analysis of Microarrays identified overexpression of two genes among the 4 tumors with 6p21 amplifications. Each point represents the observed t -statistic, plotted against its expected score from permutation tests. The most significant gene is VEGFA , followed by TMEM63B (indicated in purple). The dashed lines indicate the confidence interval for the random, null distribution. (B) VEGFA or (C) TMEM63B is overexpressed in tumors with high-level gains of 6p21. Each of the 91 points represents the corresponding copy number and expression level for a single tumor. The horizontal axis indicates the median inferred copy number for 68 single nucleotide polymorphism probes in the minimal overlapping region at 6p21. The vertical axis indicates the log2 ratio between the expression level of each tumor, compared to the median of 10 normal liver samples on the Affymetrix U133 Plus 2 array. (D) Higher VEGFA expression is associated with increased copy numbers in an independent cohort. Each of the 45 points represents a single tumor chosen from the panel of 210 tumors evaluated by fluorescence in situ hybridization. The horizontal axis indicates the average number of VEGFA probes per nucleus. The vertical axis indicates the negative ΔΔ C t values of VEGFA normalized to ACTB , compared to the median of 5 uninvolved, cirrhotic tissue samples. Tumors with more than 4 copies of VEGFA are shown in red ( n = 10), tumors with gains of chromosome 6 are shown in black ( n = 19), and diploid tumors are shown in grey ( n = 16). The average negative ΔΔ C t value for each of these three classes is plotted with a dashed blue line.

    Techniques Used: Over Expression, Expressing, Fluorescence, In Situ Hybridization

    6) Product Images from "Energy status and HIF signalling in chorionic villi show no evidence of hypoxic stress during human early placental development"

    Article Title: Energy status and HIF signalling in chorionic villi show no evidence of hypoxic stress during human early placental development

    Journal: Molecular Human Reproduction

    doi: 10.1093/molehr/gau105

    The effect of p38 inhibition on HIF-1α protein ( A ) and VEGFA and GLUT3 mRNA ( B ) in first trimester placental explants subjected to 2 or 21% O 2 ± 1 mM H 2 O 2 for 6 h. (A) Lysates from first trimester explants cultured with or without the p38 inhibitor PD169316 (p38i; 10 µM) were immunoblotted with HIF-1α antibody. Poncaeu S staining served to normalize gel loading. Normalized results (±SEM) are plotted, expressing T 0 samples as 100%. (B) RNA was isolated and relative levels of VEGFA and GLUT3 mRNA were detected using quantitative real-time RT–PCR. VEGFA and GLUT3 mRNA levels were normalized to the 18S RNA levels. Different letters indicate groups that are significantly different ( P
    Figure Legend Snippet: The effect of p38 inhibition on HIF-1α protein ( A ) and VEGFA and GLUT3 mRNA ( B ) in first trimester placental explants subjected to 2 or 21% O 2 ± 1 mM H 2 O 2 for 6 h. (A) Lysates from first trimester explants cultured with or without the p38 inhibitor PD169316 (p38i; 10 µM) were immunoblotted with HIF-1α antibody. Poncaeu S staining served to normalize gel loading. Normalized results (±SEM) are plotted, expressing T 0 samples as 100%. (B) RNA was isolated and relative levels of VEGFA and GLUT3 mRNA were detected using quantitative real-time RT–PCR. VEGFA and GLUT3 mRNA levels were normalized to the 18S RNA levels. Different letters indicate groups that are significantly different ( P

    Techniques Used: Inhibition, Cell Culture, Staining, Expressing, Isolation, Quantitative RT-PCR

    VEGFA and PlGF mRNA in T 0 controls ( T 0 ), and in explants cultured under 2 or 21% O 2 ± 1 mM H 2 O 2 for 6 h. RNA was isolated and relative levels of VEGFA and PlGF mRNA were detected using quantitative real-time RT–PCR. VEGFA and PlGF mRNA levels were normalized to the 18S RNA levels. Significant differences ( P
    Figure Legend Snippet: VEGFA and PlGF mRNA in T 0 controls ( T 0 ), and in explants cultured under 2 or 21% O 2 ± 1 mM H 2 O 2 for 6 h. RNA was isolated and relative levels of VEGFA and PlGF mRNA were detected using quantitative real-time RT–PCR. VEGFA and PlGF mRNA levels were normalized to the 18S RNA levels. Significant differences ( P

    Techniques Used: Cell Culture, Isolation, Quantitative RT-PCR

    Related Articles

    Real-time Polymerase Chain Reaction:

    Article Title: Constitutive activation of MET signaling impairs myogenic differentiation of rhabdomyosarcoma and promotes its development and progression
    Article Snippet: Expression of TPR-MET mRNA was evaluated by PCR with a forward primer 5′-GAGCCAATTTACAAGAACAAAGGA-3′ and reverse primer 5′-ATACTGCACTTGTCGGCATGAA-3′. .. Quantitative real-time PCR Gene expression was determined by qRT-PCR analysis on ABI PRISM 7300 Sequence Detection System (Applied Biosystems, Foster City, CA, USA) using Blank qPCR Master Mix (EURx) and the following Taq-Man probes (Applied Biosystems): human: GAPDH (Hs99999905_m1), MET (Hs01565589_m1), VEGF (Hs00173626_m1), MMP9 (Hs00234579_m1), MYF5 (Hs00271574_m1), MYOD (Hs00159528_m1), MRF4 (Hs01547104_g1), MEF2A (Hs01050409_m1), MYOSTATIN (Hs00976237_m1), MYOGENIN (Hs01032275_m1), MYH2 (Hs00430042_m1), PAX3 (Hs00240950_m1), PAX7 (Hs00242962_m1), SNAI1 (Hs00195591_m1), RUNX2 (Hs00231692_m1), PPARG2 (Hs01115513_m1) and mouse: GAPDH (Mm99999915_g1). .. For evaluation of miRNA expression by quantitative real-time PCR Sybr Green qPCR Master MIX (EURx) and universal reverse primer from NCode VILO miRNA cDNA Synthesis Kit (Invitrogen) were used with the following forward primers: U6 snRNA: 5-CGCAAGGATGACACGCAAA TTC-3′ miR-1: 5′-GCTGGAATGTAAAGAAGTATGT ATAA-3′ miR-206: 5-TGGAATGTAAGGAAGTGTGTGG-3′ miR-133a-5p: 5-GCAGCTGGTAAAATGGAACCA AAT-3′ miR-133a-3p: 5′-TGGTCCCCTTCAACCAGCTG-3′ miR-133b: 5′-TTTGGTCCCCTTCAACCAGCTA-3′ miR-378a-5p: 5′-CCTGACTCCAGGTCCTGTGT-3′ miR-378a-3p: 5′-ACTGGACTTGGAGTCAG AAGG-3′ The mRNA expression level for all samples was normalized to the housekeeping gene GAPDH, whereas miRNA level was normalized to the housekeeping U6 snRNA level.

    Expressing:

    Article Title: Constitutive activation of MET signaling impairs myogenic differentiation of rhabdomyosarcoma and promotes its development and progression
    Article Snippet: Expression of TPR-MET mRNA was evaluated by PCR with a forward primer 5′-GAGCCAATTTACAAGAACAAAGGA-3′ and reverse primer 5′-ATACTGCACTTGTCGGCATGAA-3′. .. Quantitative real-time PCR Gene expression was determined by qRT-PCR analysis on ABI PRISM 7300 Sequence Detection System (Applied Biosystems, Foster City, CA, USA) using Blank qPCR Master Mix (EURx) and the following Taq-Man probes (Applied Biosystems): human: GAPDH (Hs99999905_m1), MET (Hs01565589_m1), VEGF (Hs00173626_m1), MMP9 (Hs00234579_m1), MYF5 (Hs00271574_m1), MYOD (Hs00159528_m1), MRF4 (Hs01547104_g1), MEF2A (Hs01050409_m1), MYOSTATIN (Hs00976237_m1), MYOGENIN (Hs01032275_m1), MYH2 (Hs00430042_m1), PAX3 (Hs00240950_m1), PAX7 (Hs00242962_m1), SNAI1 (Hs00195591_m1), RUNX2 (Hs00231692_m1), PPARG2 (Hs01115513_m1) and mouse: GAPDH (Mm99999915_g1). .. For evaluation of miRNA expression by quantitative real-time PCR Sybr Green qPCR Master MIX (EURx) and universal reverse primer from NCode VILO miRNA cDNA Synthesis Kit (Invitrogen) were used with the following forward primers: U6 snRNA: 5-CGCAAGGATGACACGCAAA TTC-3′ miR-1: 5′-GCTGGAATGTAAAGAAGTATGT ATAA-3′ miR-206: 5-TGGAATGTAAGGAAGTGTGTGG-3′ miR-133a-5p: 5-GCAGCTGGTAAAATGGAACCA AAT-3′ miR-133a-3p: 5′-TGGTCCCCTTCAACCAGCTG-3′ miR-133b: 5′-TTTGGTCCCCTTCAACCAGCTA-3′ miR-378a-5p: 5′-CCTGACTCCAGGTCCTGTGT-3′ miR-378a-3p: 5′-ACTGGACTTGGAGTCAG AAGG-3′ The mRNA expression level for all samples was normalized to the housekeeping gene GAPDH, whereas miRNA level was normalized to the housekeeping U6 snRNA level.

    Article Title: Focal Gains of Vascular Endothelial Growth Factor A and Molecular Classification of Hepatocellular Carcinoma
    Article Snippet: Total RNA was extracted from 3 to 4 tissue sections, each 10 microns thick, using the TRIzol LS reagent (Invitrogen). cDNA synthesis and PCR conditions were conducted as previously described ( ). .. Expression levels were measured with Taqman Probes® Hs00173626-m1 for VEGFA and Hs03023943-g1 for ACTB obtained from Taqman Gene Expression Assays® (Applied Biosystems). ..

    Article Title: Kaposi's Sarcoma Herpesvirus K15 Protein Contributes to Virus-Induced Angiogenesis by Recruiting PLC?1 and Activating NFAT1-dependent RCAN1 Expression
    Article Snippet: Amplification was performed in 10 µl reactions with TaqMan Universal PCR Master Mix under recommended conditions (Applied Biosystems; #4364341). .. The following TaqMan gene expression assays (Applied Biosystems: #4331182) were used: Hs01120954_m1 (RCAN1); Hs00173626_m1 (VEGFA); Hs99999905_m1 (GAPDH). .. To detect K15 mRNA, primers 5′-CGGAAGAATCACGTGAAC-3′ (sense) and 5′-CGGTGTCTATACGGAAGG-3′ (antisense) and a dually labeled probe 5′-FAM-TCACCACAGCCAGACCAATCA-TAMRA-3′ were used.

    Quantitative RT-PCR:

    Article Title: Constitutive activation of MET signaling impairs myogenic differentiation of rhabdomyosarcoma and promotes its development and progression
    Article Snippet: Expression of TPR-MET mRNA was evaluated by PCR with a forward primer 5′-GAGCCAATTTACAAGAACAAAGGA-3′ and reverse primer 5′-ATACTGCACTTGTCGGCATGAA-3′. .. Quantitative real-time PCR Gene expression was determined by qRT-PCR analysis on ABI PRISM 7300 Sequence Detection System (Applied Biosystems, Foster City, CA, USA) using Blank qPCR Master Mix (EURx) and the following Taq-Man probes (Applied Biosystems): human: GAPDH (Hs99999905_m1), MET (Hs01565589_m1), VEGF (Hs00173626_m1), MMP9 (Hs00234579_m1), MYF5 (Hs00271574_m1), MYOD (Hs00159528_m1), MRF4 (Hs01547104_g1), MEF2A (Hs01050409_m1), MYOSTATIN (Hs00976237_m1), MYOGENIN (Hs01032275_m1), MYH2 (Hs00430042_m1), PAX3 (Hs00240950_m1), PAX7 (Hs00242962_m1), SNAI1 (Hs00195591_m1), RUNX2 (Hs00231692_m1), PPARG2 (Hs01115513_m1) and mouse: GAPDH (Mm99999915_g1). .. For evaluation of miRNA expression by quantitative real-time PCR Sybr Green qPCR Master MIX (EURx) and universal reverse primer from NCode VILO miRNA cDNA Synthesis Kit (Invitrogen) were used with the following forward primers: U6 snRNA: 5-CGCAAGGATGACACGCAAA TTC-3′ miR-1: 5′-GCTGGAATGTAAAGAAGTATGT ATAA-3′ miR-206: 5-TGGAATGTAAGGAAGTGTGTGG-3′ miR-133a-5p: 5-GCAGCTGGTAAAATGGAACCA AAT-3′ miR-133a-3p: 5′-TGGTCCCCTTCAACCAGCTG-3′ miR-133b: 5′-TTTGGTCCCCTTCAACCAGCTA-3′ miR-378a-5p: 5′-CCTGACTCCAGGTCCTGTGT-3′ miR-378a-3p: 5′-ACTGGACTTGGAGTCAG AAGG-3′ The mRNA expression level for all samples was normalized to the housekeeping gene GAPDH, whereas miRNA level was normalized to the housekeeping U6 snRNA level.

    Sequencing:

    Article Title: Constitutive activation of MET signaling impairs myogenic differentiation of rhabdomyosarcoma and promotes its development and progression
    Article Snippet: Expression of TPR-MET mRNA was evaluated by PCR with a forward primer 5′-GAGCCAATTTACAAGAACAAAGGA-3′ and reverse primer 5′-ATACTGCACTTGTCGGCATGAA-3′. .. Quantitative real-time PCR Gene expression was determined by qRT-PCR analysis on ABI PRISM 7300 Sequence Detection System (Applied Biosystems, Foster City, CA, USA) using Blank qPCR Master Mix (EURx) and the following Taq-Man probes (Applied Biosystems): human: GAPDH (Hs99999905_m1), MET (Hs01565589_m1), VEGF (Hs00173626_m1), MMP9 (Hs00234579_m1), MYF5 (Hs00271574_m1), MYOD (Hs00159528_m1), MRF4 (Hs01547104_g1), MEF2A (Hs01050409_m1), MYOSTATIN (Hs00976237_m1), MYOGENIN (Hs01032275_m1), MYH2 (Hs00430042_m1), PAX3 (Hs00240950_m1), PAX7 (Hs00242962_m1), SNAI1 (Hs00195591_m1), RUNX2 (Hs00231692_m1), PPARG2 (Hs01115513_m1) and mouse: GAPDH (Mm99999915_g1). .. For evaluation of miRNA expression by quantitative real-time PCR Sybr Green qPCR Master MIX (EURx) and universal reverse primer from NCode VILO miRNA cDNA Synthesis Kit (Invitrogen) were used with the following forward primers: U6 snRNA: 5-CGCAAGGATGACACGCAAA TTC-3′ miR-1: 5′-GCTGGAATGTAAAGAAGTATGT ATAA-3′ miR-206: 5-TGGAATGTAAGGAAGTGTGTGG-3′ miR-133a-5p: 5-GCAGCTGGTAAAATGGAACCA AAT-3′ miR-133a-3p: 5′-TGGTCCCCTTCAACCAGCTG-3′ miR-133b: 5′-TTTGGTCCCCTTCAACCAGCTA-3′ miR-378a-5p: 5′-CCTGACTCCAGGTCCTGTGT-3′ miR-378a-3p: 5′-ACTGGACTTGGAGTCAG AAGG-3′ The mRNA expression level for all samples was normalized to the housekeeping gene GAPDH, whereas miRNA level was normalized to the housekeeping U6 snRNA level.

    Negative Control:

    Article Title: Endothelial Differentiation of Adipose Tissue-Derived Mesenchymal Stromal Cells in Glioma Tumors: Implications for Cell-Based Therapy
    Article Snippet: A pre-amplification stage was performed with TaqMan PreAmp Master Mix (Applied Biosystems). .. FAM-labeled primer/probes were purchased from Applied Biosystems: EGR-3 (Hs00231780_m1); CD31 (Hs00169777_m1); ILK (Hs00177914_m1); SD1-α (Hs00930455_m1); VEGFa1 (Hs00173626_m1); Notch1 (Hs01062014_m1); negative control Osteocalcin BGLAP (Hs01587813_g1). .. The threshold cycle (Ct) method was used to quantify relative expression for each gene using GAPDH as endogenous reference.

    Binding Assay:

    Article Title: Novel TPO receptor agonist TA-316 contributes to platelet biogenesis from human iPS cells
    Article Snippet: RT-PCR was carried out for 40 to 50 cycles of 1 minute at 60°C and 15 seconds at 95°C in an ABI PRISM 7700 Sequence Detector (Applied Biosystems, Foster City, CA). .. All Taqman primers and probes were obtained from Applied Biosystems: glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Hs02758991_g1); GATA binding protein 1 (GATA1; Hs01085823_m1); zinc finger protein, FOG family member 1 (ZFPM1; Hs00542350_m1); nuclear factor, erythroid 2 (NFE2; Hs00232351_m1); Fli-1 proto-oncogene ETS transcription factor (FLI1; Hs00956711_m1); von Willebrand factor (VWF; Hs04397751_m1); VEGF A (VEGFA; Hs00173626_m1); and basic FGF2 (Hs00266645_m1). ..

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    Thermo Fisher gene exp vegfa hs00173626 m1
    In vitro tube formation by hAMSC requires Notch1 expression . Phase contrast photomicrographs ( a ) of untransduced (left); αNotch1 shRNA transduced (middle) and mock ΦshRNA transduced (right) PECAM:PL-G/RL-R-tTK-hAMSCs incubated in Matrigel for 4 hours. ( b ) RNA extracted from cells in matrigel was assayed by RT-PCR to determine transcription the levels of the indicated endothelial (ILK, SDF1, EGR3,PECAM-1, <t>VEGFA-1,</t> Notch-1) and bone BGLAP/Octeocalcin differentiation markers. The histogram shows % fold change relative to the control un-transduced PECAM:PL-G/RL-R-tTK-hAMSCs. * P
    Gene Exp Vegfa Hs00173626 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    In vitro tube formation by hAMSC requires Notch1 expression . Phase contrast photomicrographs ( a ) of untransduced (left); αNotch1 shRNA transduced (middle) and mock ΦshRNA transduced (right) PECAM:PL-G/RL-R-tTK-hAMSCs incubated in Matrigel for 4 hours. ( b ) RNA extracted from cells in matrigel was assayed by RT-PCR to determine transcription the levels of the indicated endothelial (ILK, SDF1, EGR3,PECAM-1, VEGFA-1, Notch-1) and bone BGLAP/Octeocalcin differentiation markers. The histogram shows % fold change relative to the control un-transduced PECAM:PL-G/RL-R-tTK-hAMSCs. * P

    Journal: Molecular Therapy

    Article Title: Endothelial Differentiation of Adipose Tissue-Derived Mesenchymal Stromal Cells in Glioma Tumors: Implications for Cell-Based Therapy

    doi: 10.1038/mt.2013.145

    Figure Lengend Snippet: In vitro tube formation by hAMSC requires Notch1 expression . Phase contrast photomicrographs ( a ) of untransduced (left); αNotch1 shRNA transduced (middle) and mock ΦshRNA transduced (right) PECAM:PL-G/RL-R-tTK-hAMSCs incubated in Matrigel for 4 hours. ( b ) RNA extracted from cells in matrigel was assayed by RT-PCR to determine transcription the levels of the indicated endothelial (ILK, SDF1, EGR3,PECAM-1, VEGFA-1, Notch-1) and bone BGLAP/Octeocalcin differentiation markers. The histogram shows % fold change relative to the control un-transduced PECAM:PL-G/RL-R-tTK-hAMSCs. * P

    Article Snippet: FAM-labeled primer/probes were purchased from Applied Biosystems: EGR-3 (Hs00231780_m1); CD31 (Hs00169777_m1); ILK (Hs00177914_m1); SD1-α (Hs00930455_m1); VEGFa1 (Hs00173626_m1); Notch1 (Hs01062014_m1); negative control Osteocalcin BGLAP (Hs01587813_g1).

    Techniques: In Vitro, Expressing, shRNA, Incubation, Reverse Transcription Polymerase Chain Reaction

    An inhibitor of FGF or VEGF receptors eliminates the superior effect of TA-316 on imMKCL proliferation. imMKCLs cultured with rhTPO (50 ng/mL), TA-316 (200 ng/mL), eltrombopag (1000 ng/mL), or DMSO (0.05%). (A) Cells were collected on day 11 (genes on). mRNA expressions of VEGFA and FGF2 (genes on) were evaluated using Taqman PCR assays. GAPDH served as an internal control. (B) FGFR antagonist PD173074 (100 nM; WAKO), VEGFR antagonist axitinib (5 nM; Toronto Research Chemicals, Toronto, Canada), or epithelial growth factor receptor antagonist gefitinib (1 μM, Santa Cruz Biotechnology, Dallas, TX) was added to the imMKCLs, which were cultured with TA-316 (200 ng/mL) for 4 days (genes on). Cell proliferation was assessed based on 0.4% Trypan blue staining (Thermo Fisher Scientific). Data represent the mean and SD; * P

    Journal: Blood Advances

    Article Title: Novel TPO receptor agonist TA-316 contributes to platelet biogenesis from human iPS cells

    doi: 10.1182/bloodadvances.2016000844

    Figure Lengend Snippet: An inhibitor of FGF or VEGF receptors eliminates the superior effect of TA-316 on imMKCL proliferation. imMKCLs cultured with rhTPO (50 ng/mL), TA-316 (200 ng/mL), eltrombopag (1000 ng/mL), or DMSO (0.05%). (A) Cells were collected on day 11 (genes on). mRNA expressions of VEGFA and FGF2 (genes on) were evaluated using Taqman PCR assays. GAPDH served as an internal control. (B) FGFR antagonist PD173074 (100 nM; WAKO), VEGFR antagonist axitinib (5 nM; Toronto Research Chemicals, Toronto, Canada), or epithelial growth factor receptor antagonist gefitinib (1 μM, Santa Cruz Biotechnology, Dallas, TX) was added to the imMKCLs, which were cultured with TA-316 (200 ng/mL) for 4 days (genes on). Cell proliferation was assessed based on 0.4% Trypan blue staining (Thermo Fisher Scientific). Data represent the mean and SD; * P

    Article Snippet: All Taqman primers and probes were obtained from Applied Biosystems: glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Hs02758991_g1); GATA binding protein 1 (GATA1; Hs01085823_m1); zinc finger protein, FOG family member 1 (ZFPM1; Hs00542350_m1); nuclear factor, erythroid 2 (NFE2; Hs00232351_m1); Fli-1 proto-oncogene ETS transcription factor (FLI1; Hs00956711_m1); von Willebrand factor (VWF; Hs04397751_m1); VEGF A (VEGFA; Hs00173626_m1); and basic FGF2 (Hs00266645_m1).

    Techniques: Cell Culture, Polymerase Chain Reaction, Staining

    K15 dependent cellular gene expression in HUVECs. (A) Microarray analysis of cellular mRNAs upregulated in HUVECs by K15 overexpression or after KSHV infection. HUVECs were transduced with a K15-expression vector (lanes 1–2) or infected with either KSHVwt or KSHVΔK15 (lanes 3–4). Depicted are the top-ranking 50 transcripts most strongly upregulated by K15 overexpression, showing an at least two-fold induction by K15 overexpression in each of two experiments performed using cells from two healthy donors (lanes 1–2). Asterisks next to fold change values within heatmap lanes 3–4 indicate genes that were induced more than two-fold in KSHVwt-infected cells compared to cells mock infected with heat-inactivated virus. Empty cells in the heatmap correspond to undetectable mRNA levels in both of the two samples compared in each lane. Arrows indicate cellular genes, inducible upon KSHVwt infection with more than two-fold elevated expression in KSHVwt- relative to KSHVΔK15-infected cells in each of the two experiments performed. (B) Quantitative PCR analysis of K15 transcript in mRNA derived from KSHVwt- or KSHVΔK15-infected cells used for the microarray experiment. (C) Quantitative PCR analysis of RCAN1/DSCR1 and VEGFA transcripts in HUVECs transduced with a retroviral K15 expression vector or with an empty control vector. (D) HUVECs from two healthy donors (Exp1–2) were infected with heat-inactivated, KSHVwt, or KSHVΔK15 and were lysed 4 days after infection. RNA was extracted, reversely transcribed and subjected to quantitative TaqMan-based PCR analysis. mRNA fold change values of KSHVwt-infected versus mock-infected (with heat-inactivated KSHVwt = HI) cells were depicted in black, values of KSHVΔK15- infected versus mock-infected cells were depicted in red. Input mRNA samples are identical to samples used in microarray experiments (compare panel A lanes 3–4).

    Journal: PLoS Pathogens

    Article Title: Kaposi's Sarcoma Herpesvirus K15 Protein Contributes to Virus-Induced Angiogenesis by Recruiting PLC?1 and Activating NFAT1-dependent RCAN1 Expression

    doi: 10.1371/journal.ppat.1002927

    Figure Lengend Snippet: K15 dependent cellular gene expression in HUVECs. (A) Microarray analysis of cellular mRNAs upregulated in HUVECs by K15 overexpression or after KSHV infection. HUVECs were transduced with a K15-expression vector (lanes 1–2) or infected with either KSHVwt or KSHVΔK15 (lanes 3–4). Depicted are the top-ranking 50 transcripts most strongly upregulated by K15 overexpression, showing an at least two-fold induction by K15 overexpression in each of two experiments performed using cells from two healthy donors (lanes 1–2). Asterisks next to fold change values within heatmap lanes 3–4 indicate genes that were induced more than two-fold in KSHVwt-infected cells compared to cells mock infected with heat-inactivated virus. Empty cells in the heatmap correspond to undetectable mRNA levels in both of the two samples compared in each lane. Arrows indicate cellular genes, inducible upon KSHVwt infection with more than two-fold elevated expression in KSHVwt- relative to KSHVΔK15-infected cells in each of the two experiments performed. (B) Quantitative PCR analysis of K15 transcript in mRNA derived from KSHVwt- or KSHVΔK15-infected cells used for the microarray experiment. (C) Quantitative PCR analysis of RCAN1/DSCR1 and VEGFA transcripts in HUVECs transduced with a retroviral K15 expression vector or with an empty control vector. (D) HUVECs from two healthy donors (Exp1–2) were infected with heat-inactivated, KSHVwt, or KSHVΔK15 and were lysed 4 days after infection. RNA was extracted, reversely transcribed and subjected to quantitative TaqMan-based PCR analysis. mRNA fold change values of KSHVwt-infected versus mock-infected (with heat-inactivated KSHVwt = HI) cells were depicted in black, values of KSHVΔK15- infected versus mock-infected cells were depicted in red. Input mRNA samples are identical to samples used in microarray experiments (compare panel A lanes 3–4).

    Article Snippet: The following TaqMan gene expression assays (Applied Biosystems: #4331182) were used: Hs01120954_m1 (RCAN1); Hs00173626_m1 (VEGFA); Hs99999905_m1 (GAPDH).

    Techniques: Expressing, Microarray, Over Expression, Infection, Transduction, Plasmid Preparation, Real-time Polymerase Chain Reaction, Derivative Assay, Polymerase Chain Reaction

    Quantitative polymerase chain reaction was used to analyze proangiogenic gene expression by MSCs on various scaffolds. (A) VEGFA , (B) PDGF , (C) FGF1 , and (D) FGF2 expression were normalized to RPL13 expression and data were represented as ΔCt.

    Journal: Tissue Engineering. Part A

    Article Title: Bioceramic-Mediated Trophic Factor Secretion by Mesenchymal Stem Cells Enhances In Vitro Endothelial Cell Persistence and In Vivo Angiogenesis

    doi: 10.1089/ten.tea.2011.0127

    Figure Lengend Snippet: Quantitative polymerase chain reaction was used to analyze proangiogenic gene expression by MSCs on various scaffolds. (A) VEGFA , (B) PDGF , (C) FGF1 , and (D) FGF2 expression were normalized to RPL13 expression and data were represented as ΔCt.

    Article Snippet: Primers and probes for VEGFA (Hs00173626_m1), PDGF (Hs00234042_m1), FGF1 (Hs00265254_m1), and FGF2 (Hs00266645_m1) were purchased from Applied Biosystems.

    Techniques: Real-time Polymerase Chain Reaction, Expressing