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  • 90
    Thermo Fisher pcr buffer
    Pcr Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1144 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche polymerase chain reaction qrt pcr analysis
    ANG regulates primitive hematopoietic cell properties through PLXNB2 (A) Quantification of HSPC cell cycle status of Mx1-specific Plxnb2 −/− mice (n=9). (B) Serial re-plating colony assay of whole BM cells (n=2). ANG was added at each re-plating. (C) Experimental schema of transplant using Mx1-specific Plxnb2 −/− whole BM as donors. (D) Multi-lineage donor cell chimerism after competitive transplant of Mx1-specific Plxnb2 −/− donors (n=5). (E-F) ANG (0.3 μg/ml) regulates LT-HSC proliferation (E, n=3) and pro-self-renewal transcripts (F, n=3) from WT but not from Mx1-specific Plxnb2 −/− mice. (G) Quantification of HSPC cell cycle status of mAb17-treated WT or Ang −/− mice (n=6). (H-I) Cell density on day 7 (H, n=3) and <t>qRT-PCR</t> analysis of pro-self-renewal transcripts (I, n=3) from sorted WT or Ang −/− LT-HSC cultured in the presence of 0.3 μg/ml ANG, 50 μg/ml mAb17, or both. (J-K) Cell density on day 7 (J, n=3) or qRT-PCR analysis of pro-self-renewal transcripts (K, n=3) from sorted WT or Ang −/− LT-HSC cultured in the presence of 0.3 μg/ml Sema4C, 50 μg/ml mAb17, or both. (L) Experimental schema of transplant using sorted WT LT-HSC that had been cultured with 0.3 μg/ml ANG, 50 μg/ml mAb17, or both for 7 days. (M) Multi-lineage donor cell chimerism after competitive transplant of ANG and/or mAb17-treated WT LT-HSC donors (n=6). .
    Polymerase Chain Reaction Qrt Pcr Analysis, supplied by Roche, used in various techniques. Bioz Stars score: 85/100, based on 307 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche real time polymerase chain reaction qrt pcr analysis
    Transcript levels of cbs are up-regulated by 1,25(OH) 2 D 3 in murine MC3T3-E1 and primary osteoblasts. (A) Transcript levels were determined by <t>qRT-PCR</t> at different time points after treatment with 1,25(OH) 2 D 3 (10 −8 M). Each data point represents
    Real Time Polymerase Chain Reaction Qrt Pcr Analysis, supplied by Roche, used in various techniques. Bioz Stars score: 84/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche quantitative reverse transcription polymerase chain reaction qrt pcr analysis
    Transcript levels of cbs are up-regulated by 1,25(OH) 2 D 3 in murine MC3T3-E1 and primary osteoblasts. (A) Transcript levels were determined by <t>qRT-PCR</t> at different time points after treatment with 1,25(OH) 2 D 3 (10 −8 M). Each data point represents
    Quantitative Reverse Transcription Polymerase Chain Reaction Qrt Pcr Analysis, supplied by Roche, used in various techniques. Bioz Stars score: 87/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche quantitative real time polymerase chain reaction qrt pcr analysis
    Transcript levels of cbs are up-regulated by 1,25(OH) 2 D 3 in murine MC3T3-E1 and primary osteoblasts. (A) Transcript levels were determined by <t>qRT-PCR</t> at different time points after treatment with 1,25(OH) 2 D 3 (10 −8 M). Each data point represents
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr Analysis, supplied by Roche, used in various techniques. Bioz Stars score: 85/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche qrt pcr tripure reagent
    Transcript levels of cbs are up-regulated by 1,25(OH) 2 D 3 in murine MC3T3-E1 and primary osteoblasts. (A) Transcript levels were determined by <t>qRT-PCR</t> at different time points after treatment with 1,25(OH) 2 D 3 (10 −8 M). Each data point represents
    Qrt Pcr Tripure Reagent, supplied by Roche, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche qrt pcr qrt pcr
    Expression profiles of loquat sucrose synthase and sucrose phosphate synthase ( EjSPS and EjSS ) using <t>qRT-PCR</t> during fruit development. Each point is the mean of three determinations. Vertical bars, if larger than symbols, represent the standard error of the mean ( n = 3).
    Qrt Pcr Qrt Pcr, supplied by Roche, used in various techniques. Bioz Stars score: 91/100, based on 440 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche qrt pcr total rna
    Effect of N. arvensis extracts in IB3-1 cells. (A) Effect of N. arvensis extracts on IL-8 mRNA expression in IB3-1 cells. IB3-1 cells were treated with the chloroform extract (solved in EtOH/DMSO 95/5) (A) at different concentrations (0.1–200 μg/ml) for 16 h before infection with PAO1 (100 CFU/cell) for further 4 h. IL-8 mRNA expression was quantified by <t>qRT-PCR.</t> Expression of IL-8 mRNA was measured by Real-Time qPCR and obtained by comparing the ratio IL-8 and the housekeeping gene GAPDH between non-infected and infected cells. The results are expressed as the % of untreated cells. Data are mean ±SEM of three independent experiments performed in duplicate. Dashed line corresponds to cells treated with solvent alone. (B) Effect of N. arvensis extract in PAO1 infected IB3-1 cells after different incubation times. The N. arvensis extract (10 μg/ml) was added to IB3-1 cells 24, 4, and 2 h before, simultaneously or 2 h post PAO1 infection (100 CFU/cell). IL-8 mRNA expression was measured as indicated in (A) . Data are mean ±SEM of three independent experiments performed in duplicate. Dashed line corresponds to cells treated with solvent alone. (C) PAO1 growth. Bacteria were cultured overnight at 37°C in the presence of solvent or ranging doses (0.1–1000 μg/ml) of N. arvensis extract. Bacterial growth was monitored by absorbance measures at 660 nm. A representative experiment performed in duplicate is shown. (D) Adhesion of PAO1 to IB3.1 cells. 500,000 IB3-1 cells on Petri dishes, in duplicate, were treated for 24 h with 10 μg/ml N. arvensis extract. Different amounts of 35 S-PAO1, expressed as CFU/well, were added to the wells and incubated as described in section “Materials and Methods”. Data reported in the figure are the specific binding calculated by subtracting counts obtained in the presence of 100-fold excess of non-labeled PAO1 and are expressed as CFU/well. Data are mean ±SEM of three independent experiments performed in duplicate. (E) Effects of N. arvensis extracts on cell growth in IB3-1 cells. IB3-1 cells were incubated with increasing concentrations (1–200 μg/ml) of the chloroform extract (solved in EtOH/DMSO) of N. arvensis for 24 and 48 h. Cell viability was measured by cytometer analysis. Data are expressed as % control (solvent) and are relative to a representative experiment performed in duplicate. Dashed line corresponds to cells treated with solvent alone.
    Qrt Pcr Total Rna, supplied by Roche, used in various techniques. Bioz Stars score: 91/100, based on 200 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche quantitative reverse transcription pcr qrt pcr analysis
    Spatial regulation of SA accumulation leading to an organized multicellular response in ETI. (A) Site-specific sampling for SA and PR1 analyses at 7 h.p.i. A nearly complete half of a pPR1-YFP-NLS leaf was fully infiltrated with Pst_a2 (OD 600 = 0.2, upper) or 10 mM MgCl 2 (mock, lower). The leaf was divided into four areas along the mid-rib (numbered 1–4), and three leaf disks (2 mm in diameter) per area were sampled, as shown in a dashed ellipse for zone 1 in the upper right pictures. Representative sample pictures are shown. Scale bars = 2.5 mm. (B) Intensity profiles of YFP and autofluorescence in the white boxes in the Pst_a2 -treated leaf in (A) along the red arrow. (C) Intensity profiles of YFP and autofluorescence in the white boxes in the mock-treated leaf in (A) along the red arrow. (D) The endogenous PR1 expression levels in the four zones were measured by <t>qRT–PCR.</t> Eighteen leaf disks, corresponding to one zone, from six leaves were pooled as one sample. Bars represent means � SD of three biological replicates. (E) The free SA and SA glycoside (SAG) levels in the four zones. Three disks from one zone from one leaf were pooled and analyzed. Bars represent means � SD of three leaves. Experiments were repeated twice with similar results. (F) A schematic summary of an organized concentric pattern of the inner SA and the outer JA active domains which appeared around the infection site of Pst_a2 . (See also Supplementary Fig. S4 ).
    Quantitative Reverse Transcription Pcr Qrt Pcr Analysis, supplied by Roche, used in various techniques. Bioz Stars score: 80/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche lightcycler 96 qrt pcr software version 1 1 0 1320
    Spatial regulation of SA accumulation leading to an organized multicellular response in ETI. (A) Site-specific sampling for SA and PR1 analyses at 7 h.p.i. A nearly complete half of a pPR1-YFP-NLS leaf was fully infiltrated with Pst_a2 (OD 600 = 0.2, upper) or 10 mM MgCl 2 (mock, lower). The leaf was divided into four areas along the mid-rib (numbered 1–4), and three leaf disks (2 mm in diameter) per area were sampled, as shown in a dashed ellipse for zone 1 in the upper right pictures. Representative sample pictures are shown. Scale bars = 2.5 mm. (B) Intensity profiles of YFP and autofluorescence in the white boxes in the Pst_a2 -treated leaf in (A) along the red arrow. (C) Intensity profiles of YFP and autofluorescence in the white boxes in the mock-treated leaf in (A) along the red arrow. (D) The endogenous PR1 expression levels in the four zones were measured by <t>qRT–PCR.</t> Eighteen leaf disks, corresponding to one zone, from six leaves were pooled as one sample. Bars represent means � SD of three biological replicates. (E) The free SA and SA glycoside (SAG) levels in the four zones. Three disks from one zone from one leaf were pooled and analyzed. Bars represent means � SD of three leaves. Experiments were repeated twice with similar results. (F) A schematic summary of an organized concentric pattern of the inner SA and the outer JA active domains which appeared around the infection site of Pst_a2 . (See also Supplementary Fig. S4 ).
    Lightcycler 96 Qrt Pcr Software Version 1 1 0 1320, supplied by Roche, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche lightcycler 480 sybr green i master qrt pcr kit
    Spatial regulation of SA accumulation leading to an organized multicellular response in ETI. (A) Site-specific sampling for SA and PR1 analyses at 7 h.p.i. A nearly complete half of a pPR1-YFP-NLS leaf was fully infiltrated with Pst_a2 (OD 600 = 0.2, upper) or 10 mM MgCl 2 (mock, lower). The leaf was divided into four areas along the mid-rib (numbered 1–4), and three leaf disks (2 mm in diameter) per area were sampled, as shown in a dashed ellipse for zone 1 in the upper right pictures. Representative sample pictures are shown. Scale bars = 2.5 mm. (B) Intensity profiles of YFP and autofluorescence in the white boxes in the Pst_a2 -treated leaf in (A) along the red arrow. (C) Intensity profiles of YFP and autofluorescence in the white boxes in the mock-treated leaf in (A) along the red arrow. (D) The endogenous PR1 expression levels in the four zones were measured by <t>qRT–PCR.</t> Eighteen leaf disks, corresponding to one zone, from six leaves were pooled as one sample. Bars represent means � SD of three biological replicates. (E) The free SA and SA glycoside (SAG) levels in the four zones. Three disks from one zone from one leaf were pooled and analyzed. Bars represent means � SD of three leaves. Experiments were repeated twice with similar results. (F) A schematic summary of an organized concentric pattern of the inner SA and the outer JA active domains which appeared around the infection site of Pst_a2 . (See also Supplementary Fig. S4 ).
    Lightcycler 480 Sybr Green I Master Qrt Pcr Kit, supplied by Roche, used in various techniques. Bioz Stars score: 83/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher superscript iii platinum one step rt pcr kit
    Spatial regulation of SA accumulation leading to an organized multicellular response in ETI. (A) Site-specific sampling for SA and PR1 analyses at 7 h.p.i. A nearly complete half of a pPR1-YFP-NLS leaf was fully infiltrated with Pst_a2 (OD 600 = 0.2, upper) or 10 mM MgCl 2 (mock, lower). The leaf was divided into four areas along the mid-rib (numbered 1–4), and three leaf disks (2 mm in diameter) per area were sampled, as shown in a dashed ellipse for zone 1 in the upper right pictures. Representative sample pictures are shown. Scale bars = 2.5 mm. (B) Intensity profiles of YFP and autofluorescence in the white boxes in the Pst_a2 -treated leaf in (A) along the red arrow. (C) Intensity profiles of YFP and autofluorescence in the white boxes in the mock-treated leaf in (A) along the red arrow. (D) The endogenous PR1 expression levels in the four zones were measured by <t>qRT–PCR.</t> Eighteen leaf disks, corresponding to one zone, from six leaves were pooled as one sample. Bars represent means � SD of three biological replicates. (E) The free SA and SA glycoside (SAG) levels in the four zones. Three disks from one zone from one leaf were pooled and analyzed. Bars represent means � SD of three leaves. Experiments were repeated twice with similar results. (F) A schematic summary of an organized concentric pattern of the inner SA and the outer JA active domains which appeared around the infection site of Pst_a2 . (See also Supplementary Fig. S4 ).
    Superscript Iii Platinum One Step Rt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche reverse transcriptase pcr qrt pcr lightcycler 480ii
    Spatial regulation of SA accumulation leading to an organized multicellular response in ETI. (A) Site-specific sampling for SA and PR1 analyses at 7 h.p.i. A nearly complete half of a pPR1-YFP-NLS leaf was fully infiltrated with Pst_a2 (OD 600 = 0.2, upper) or 10 mM MgCl 2 (mock, lower). The leaf was divided into four areas along the mid-rib (numbered 1–4), and three leaf disks (2 mm in diameter) per area were sampled, as shown in a dashed ellipse for zone 1 in the upper right pictures. Representative sample pictures are shown. Scale bars = 2.5 mm. (B) Intensity profiles of YFP and autofluorescence in the white boxes in the Pst_a2 -treated leaf in (A) along the red arrow. (C) Intensity profiles of YFP and autofluorescence in the white boxes in the mock-treated leaf in (A) along the red arrow. (D) The endogenous PR1 expression levels in the four zones were measured by <t>qRT–PCR.</t> Eighteen leaf disks, corresponding to one zone, from six leaves were pooled as one sample. Bars represent means � SD of three biological replicates. (E) The free SA and SA glycoside (SAG) levels in the four zones. Three disks from one zone from one leaf were pooled and analyzed. Bars represent means � SD of three leaves. Experiments were repeated twice with similar results. (F) A schematic summary of an organized concentric pattern of the inner SA and the outer JA active domains which appeared around the infection site of Pst_a2 . (See also Supplementary Fig. S4 ).
    Reverse Transcriptase Pcr Qrt Pcr Lightcycler 480ii, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche quantitative rt pcr qrt pcr qrt pcr analysis
    MCF10A cells expressing DCLK1 harbor abnormal chromosomes. A: Whole cell lysates (60 μg of total protein) of parent MCF10A, MCF10A-DCLK1 and MCF10A-DCLK1-K419R cells were immunoblotted by the indicated antibodies. B and C: cDNAs were prepared from parent MCF10A and MCF10A-DCLK1 cells and <t>qRT-PCR</t> was performed for indicated genes. The data were normalized against GAPDH and are shown as mean +/- SD. n.s., not significant; * *, p
    Quantitative Rt Pcr Qrt Pcr Qrt Pcr Analysis, supplied by Roche, used in various techniques. Bioz Stars score: 82/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche pcr quantitative real time pcr qrt pcr
    si-cJun and sh-cJun activity in melanoma after AGO2 re-expression. ( A ) <t>cJun-qRT–PCR</t> analysis of Mel-Ju cells after 2 μ g pAGO2 or 2 μ g pIRES (mock) transfection and si-cJun treatment (cJun inhibition per pmol si-cJun). cJun inhibition increases after AGO2 re-expression up to 70% compared with mock. ( B ) cJun qRT–PCR (* P
    Pcr Quantitative Real Time Pcr Qrt Pcr, supplied by Roche, used in various techniques. Bioz Stars score: 77/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche quantitative rt pcr quantitative rt pcr qrt pcr analyses
    si-cJun and sh-cJun activity in melanoma after AGO2 re-expression. ( A ) <t>cJun-qRT–PCR</t> analysis of Mel-Ju cells after 2 μ g pAGO2 or 2 μ g pIRES (mock) transfection and si-cJun treatment (cJun inhibition per pmol si-cJun). cJun inhibition increases after AGO2 re-expression up to 70% compared with mock. ( B ) cJun qRT–PCR (* P
    Quantitative Rt Pcr Quantitative Rt Pcr Qrt Pcr Analyses, supplied by Roche, used in various techniques. Bioz Stars score: 79/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche 480 real time pcr system
    si-cJun and sh-cJun activity in melanoma after AGO2 re-expression. ( A ) <t>cJun-qRT–PCR</t> analysis of Mel-Ju cells after 2 μ g pAGO2 or 2 μ g pIRES (mock) transfection and si-cJun treatment (cJun inhibition per pmol si-cJun). cJun inhibition increases after AGO2 re-expression up to 70% compared with mock. ( B ) cJun qRT–PCR (* P
    480 Real Time Pcr System, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 274 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche lightcycler 480 real time pcr system
    si-cJun and sh-cJun activity in melanoma after AGO2 re-expression. ( A ) <t>cJun-qRT–PCR</t> analysis of Mel-Ju cells after 2 μ g pAGO2 or 2 μ g pIRES (mock) transfection and si-cJun treatment (cJun inhibition per pmol si-cJun). cJun inhibition increases after AGO2 re-expression up to 70% compared with mock. ( B ) cJun qRT–PCR (* P
    Lightcycler 480 Real Time Pcr System, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 11420 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche rt pcr qrt pcr analysis adopted faststart universal sybr green master
    si-cJun and sh-cJun activity in melanoma after AGO2 re-expression. ( A ) <t>cJun-qRT–PCR</t> analysis of Mel-Ju cells after 2 μ g pAGO2 or 2 μ g pIRES (mock) transfection and si-cJun treatment (cJun inhibition per pmol si-cJun). cJun inhibition increases after AGO2 re-expression up to 70% compared with mock. ( B ) cJun qRT–PCR (* P
    Rt Pcr Qrt Pcr Analysis Adopted Faststart Universal Sybr Green Master, supplied by Roche, used in various techniques. Bioz Stars score: 81/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche light cycler 480 sequence detection system
    si-cJun and sh-cJun activity in melanoma after AGO2 re-expression. ( A ) <t>cJun-qRT–PCR</t> analysis of Mel-Ju cells after 2 μ g pAGO2 or 2 μ g pIRES (mock) transfection and si-cJun treatment (cJun inhibition per pmol si-cJun). cJun inhibition increases after AGO2 re-expression up to 70% compared with mock. ( B ) cJun qRT–PCR (* P
    Light Cycler 480 Sequence Detection System, supplied by Roche, used in various techniques. Bioz Stars score: 91/100, based on 148 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche molecular light cycler
    si-cJun and sh-cJun activity in melanoma after AGO2 re-expression. ( A ) <t>cJun-qRT–PCR</t> analysis of Mel-Ju cells after 2 μ g pAGO2 or 2 μ g pIRES (mock) transfection and si-cJun treatment (cJun inhibition per pmol si-cJun). cJun inhibition increases after AGO2 re-expression up to 70% compared with mock. ( B ) cJun qRT–PCR (* P
    Molecular Light Cycler, supplied by Roche, used in various techniques. Bioz Stars score: 84/100, based on 46 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman gene expression assays
    si-cJun and sh-cJun activity in melanoma after AGO2 re-expression. ( A ) <t>cJun-qRT–PCR</t> analysis of Mel-Ju cells after 2 μ g pAGO2 or 2 μ g pIRES (mock) transfection and si-cJun treatment (cJun inhibition per pmol si-cJun). cJun inhibition increases after AGO2 re-expression up to 70% compared with mock. ( B ) cJun qRT–PCR (* P
    Taqman Gene Expression Assays, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 37463 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche lightcycler 96 system
    si-cJun and sh-cJun activity in melanoma after AGO2 re-expression. ( A ) <t>cJun-qRT–PCR</t> analysis of Mel-Ju cells after 2 μ g pAGO2 or 2 μ g pIRES (mock) transfection and si-cJun treatment (cJun inhibition per pmol si-cJun). cJun inhibition increases after AGO2 re-expression up to 70% compared with mock. ( B ) cJun qRT–PCR (* P
    Lightcycler 96 System, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 3171 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    ANG regulates primitive hematopoietic cell properties through PLXNB2 (A) Quantification of HSPC cell cycle status of Mx1-specific Plxnb2 −/− mice (n=9). (B) Serial re-plating colony assay of whole BM cells (n=2). ANG was added at each re-plating. (C) Experimental schema of transplant using Mx1-specific Plxnb2 −/− whole BM as donors. (D) Multi-lineage donor cell chimerism after competitive transplant of Mx1-specific Plxnb2 −/− donors (n=5). (E-F) ANG (0.3 μg/ml) regulates LT-HSC proliferation (E, n=3) and pro-self-renewal transcripts (F, n=3) from WT but not from Mx1-specific Plxnb2 −/− mice. (G) Quantification of HSPC cell cycle status of mAb17-treated WT or Ang −/− mice (n=6). (H-I) Cell density on day 7 (H, n=3) and qRT-PCR analysis of pro-self-renewal transcripts (I, n=3) from sorted WT or Ang −/− LT-HSC cultured in the presence of 0.3 μg/ml ANG, 50 μg/ml mAb17, or both. (J-K) Cell density on day 7 (J, n=3) or qRT-PCR analysis of pro-self-renewal transcripts (K, n=3) from sorted WT or Ang −/− LT-HSC cultured in the presence of 0.3 μg/ml Sema4C, 50 μg/ml mAb17, or both. (L) Experimental schema of transplant using sorted WT LT-HSC that had been cultured with 0.3 μg/ml ANG, 50 μg/ml mAb17, or both for 7 days. (M) Multi-lineage donor cell chimerism after competitive transplant of ANG and/or mAb17-treated WT LT-HSC donors (n=6). .

    Journal: Cell

    Article Title: Plexin-B2 mediates physiologic and pathologic functions of angiogenin

    doi: 10.1016/j.cell.2017.10.005

    Figure Lengend Snippet: ANG regulates primitive hematopoietic cell properties through PLXNB2 (A) Quantification of HSPC cell cycle status of Mx1-specific Plxnb2 −/− mice (n=9). (B) Serial re-plating colony assay of whole BM cells (n=2). ANG was added at each re-plating. (C) Experimental schema of transplant using Mx1-specific Plxnb2 −/− whole BM as donors. (D) Multi-lineage donor cell chimerism after competitive transplant of Mx1-specific Plxnb2 −/− donors (n=5). (E-F) ANG (0.3 μg/ml) regulates LT-HSC proliferation (E, n=3) and pro-self-renewal transcripts (F, n=3) from WT but not from Mx1-specific Plxnb2 −/− mice. (G) Quantification of HSPC cell cycle status of mAb17-treated WT or Ang −/− mice (n=6). (H-I) Cell density on day 7 (H, n=3) and qRT-PCR analysis of pro-self-renewal transcripts (I, n=3) from sorted WT or Ang −/− LT-HSC cultured in the presence of 0.3 μg/ml ANG, 50 μg/ml mAb17, or both. (J-K) Cell density on day 7 (J, n=3) or qRT-PCR analysis of pro-self-renewal transcripts (K, n=3) from sorted WT or Ang −/− LT-HSC cultured in the presence of 0.3 μg/ml Sema4C, 50 μg/ml mAb17, or both. (L) Experimental schema of transplant using sorted WT LT-HSC that had been cultured with 0.3 μg/ml ANG, 50 μg/ml mAb17, or both for 7 days. (M) Multi-lineage donor cell chimerism after competitive transplant of ANG and/or mAb17-treated WT LT-HSC donors (n=6). .

    Article Snippet: M-MLV reverse transcriptase (Promega) was used to reverse transcribe total RNA into cDNA, per manufacturer’s instructions. qRT-PCR analysis was performed on a LightCycler 480 II (Roche) using SYBR Green PCR mix (Roche).

    Techniques: Mouse Assay, Colony Assay, Quantitative RT-PCR, Cell Culture

    Cell type-specific regulation of protein synthesis by ANG-PLXNB2 (A-B) Quantification of MyePro cell cycle status of WT and Mx1-specific Plxnb2 −/− (A, n=9) and mAb17-treated WT or Ang −/− mice (B, n=6). (C-D) OP-Puro incorporation following 2 h ANG treatment of WT or Mx1-specific Plxnb2 −/− HSPC and MyePro for cells in all phases (C, n=4) or in G0/G1 phase (D, n=4). (E) qRT-PCR analysis of rRNA following 2 h ANG treatment of WT or Mx1-specific Plxnb2 −/− HSPC and MyePro (n=3). (F-G) Small RNA production in Mx1-specific Plxnb2 −/− HSPC (F, n=2) or WT HSPC treated with 0.3 μg/ml ANG, 50 μg/ml mAb17, or both (G, n=2).

    Journal: Cell

    Article Title: Plexin-B2 mediates physiologic and pathologic functions of angiogenin

    doi: 10.1016/j.cell.2017.10.005

    Figure Lengend Snippet: Cell type-specific regulation of protein synthesis by ANG-PLXNB2 (A-B) Quantification of MyePro cell cycle status of WT and Mx1-specific Plxnb2 −/− (A, n=9) and mAb17-treated WT or Ang −/− mice (B, n=6). (C-D) OP-Puro incorporation following 2 h ANG treatment of WT or Mx1-specific Plxnb2 −/− HSPC and MyePro for cells in all phases (C, n=4) or in G0/G1 phase (D, n=4). (E) qRT-PCR analysis of rRNA following 2 h ANG treatment of WT or Mx1-specific Plxnb2 −/− HSPC and MyePro (n=3). (F-G) Small RNA production in Mx1-specific Plxnb2 −/− HSPC (F, n=2) or WT HSPC treated with 0.3 μg/ml ANG, 50 μg/ml mAb17, or both (G, n=2).

    Article Snippet: M-MLV reverse transcriptase (Promega) was used to reverse transcribe total RNA into cDNA, per manufacturer’s instructions. qRT-PCR analysis was performed on a LightCycler 480 II (Roche) using SYBR Green PCR mix (Roche).

    Techniques: Mouse Assay, Quantitative RT-PCR

    Verification of RNA-seq results by qRT-PCR. The DEGs in GXU-34176 vs GXU-34140 related to the category of ‘abscisic acid-activated signaling pathway’ (c71654.graph_c0 and c65832.graph_c0), ‘leaf senescence’ (c64240.graph_c0), ‘response to ethylene’ (c67492.graph_c0) and ‘cell wall modification involved in abscission’ (c65986.graph_c0). The DEGs in GN18 vs FN95–1702 related to the category of ‘response to water deprivation’ (c54647.graph_c0, c56804.graph_c0 and c61335.graph_c0) and ‘response to oxidative stress’ (c57471.graph_c0). The DEGs in GUC2 vs GUC10 related to the category of ‘ubiquitin-protein transferase activity’ (c69746.graph_c0), ‘defense response to fungus’ (c72075.graph_c0), ‘response to jasmonic acid’ (c65355.graph_c0). Data of qRT-PCR are presented as mean ± SD (n = 9) and error bars represent SD.

    Journal: Scientific Reports

    Article Title: Transcriptomic characterization and potential marker development of contrasting sugarcane cultivars

    doi: 10.1038/s41598-018-19832-x

    Figure Lengend Snippet: Verification of RNA-seq results by qRT-PCR. The DEGs in GXU-34176 vs GXU-34140 related to the category of ‘abscisic acid-activated signaling pathway’ (c71654.graph_c0 and c65832.graph_c0), ‘leaf senescence’ (c64240.graph_c0), ‘response to ethylene’ (c67492.graph_c0) and ‘cell wall modification involved in abscission’ (c65986.graph_c0). The DEGs in GN18 vs FN95–1702 related to the category of ‘response to water deprivation’ (c54647.graph_c0, c56804.graph_c0 and c61335.graph_c0) and ‘response to oxidative stress’ (c57471.graph_c0). The DEGs in GUC2 vs GUC10 related to the category of ‘ubiquitin-protein transferase activity’ (c69746.graph_c0), ‘defense response to fungus’ (c72075.graph_c0), ‘response to jasmonic acid’ (c65355.graph_c0). Data of qRT-PCR are presented as mean ± SD (n = 9) and error bars represent SD.

    Article Snippet: qRT-PCR analysis qRT-PCR was carried out to validate the reliability of differentially expressed genes in the LightCycler 480 thermocycler (Roche).

    Techniques: RNA Sequencing Assay, Quantitative RT-PCR, Modification, Activity Assay

    Tivozanib induces G2/M cell cycle arrest. ( A ) Following treatment with tivozanib for 48 h, the cell pellets were fixed and incubated with PI to analyse the cell cycle distribution on a flow cytometer. The graphs are representative of three independent experiments with similar results. ( B ) The cells were treated with tivozanib for 48 h then total RNA was harvested for qRT-PCR analysis. ( C ) Protein lysates from tivozanib-treated cells were subjected to Western blotting and probed with the indicated antibodies. β-actin was used as the loading control. The blots are representative of three independent experiments with similar outcomes. Data are given as mean ± SD. Statistically significant values of * p

    Journal: Scientific Reports

    Article Title: Blockade of vascular endothelial growth factor receptors by tivozanib has potential anti-tumour effects on human glioblastoma cells

    doi: 10.1038/srep44075

    Figure Lengend Snippet: Tivozanib induces G2/M cell cycle arrest. ( A ) Following treatment with tivozanib for 48 h, the cell pellets were fixed and incubated with PI to analyse the cell cycle distribution on a flow cytometer. The graphs are representative of three independent experiments with similar results. ( B ) The cells were treated with tivozanib for 48 h then total RNA was harvested for qRT-PCR analysis. ( C ) Protein lysates from tivozanib-treated cells were subjected to Western blotting and probed with the indicated antibodies. β-actin was used as the loading control. The blots are representative of three independent experiments with similar outcomes. Data are given as mean ± SD. Statistically significant values of * p

    Article Snippet: Analysis of gene expression by quantitative reverse transcription-PCR The quantitative reverse transcription-PCR (qRT-PCR) analysis was performed on a LightCycler® 96 instrument (Roche Molecular Diagnostics) using RealQ SYBR Green PCR reagents (Ampliqon, Copenhagen, Denmark).

    Techniques: Incubation, Flow Cytometry, Cytometry, Quantitative RT-PCR, Western Blot

    Tivozanib inhibits adhesive and invasive potential of the GBM cells. ( A ) Tivozanib-treated cells were seeded into collagen I-coated culture dishes then the adhesive cells were stained, lysed and the optical densitometry was read. ( B,C ) The effects of tivozanib on mRNA and protein levels of ICAM-1 and VCAM-1 were measured by qRT-PCR and Western blot analysis. β-actin was used as the loading control. The blots are representative of three independent experiments with similar results. ( D ) Equal amounts of secreted protein from treated cells were incubated with a synthetic substrate labelled with amino-4-trifluoromethyl coumarin (AFC). The substrate is cleaved by cathepsin B to release AFC, which is fluorometrically detected. ( E ) The conditioned media from each sample was subjected to a chromogenic substrate, which is cleaved by active uPA and produces a colorimetrically detectable product. (F) The conditioned media was collected and separated on a non-reducing polyacrylamide gel containing gelatin A. Gelatinolytic activities are visualized as clear bands against the blue background of stained gelatin. The zymograms are representative of three independent experiments with similar results. The gels were cropped and the full-length gels are presented in Supplementary Fig. 3 . (G) The cells were placed into 8-μm porous culture inserts, treated with tivozanib and allowed to migrate for 48 h. The migrated cells on the lower surface of the inserts were fixed with methanol, stained with crystal violet, lysed with 30% acetic acid and the optical densitometry was measured at 590 nm. (H) For invasion assay, the cells were placed into matrigel-coated inserts and allowed to invade through the matrigel layer for 48 h. Data are given as mean ± SD. Statistically significant values of * p

    Journal: Scientific Reports

    Article Title: Blockade of vascular endothelial growth factor receptors by tivozanib has potential anti-tumour effects on human glioblastoma cells

    doi: 10.1038/srep44075

    Figure Lengend Snippet: Tivozanib inhibits adhesive and invasive potential of the GBM cells. ( A ) Tivozanib-treated cells were seeded into collagen I-coated culture dishes then the adhesive cells were stained, lysed and the optical densitometry was read. ( B,C ) The effects of tivozanib on mRNA and protein levels of ICAM-1 and VCAM-1 were measured by qRT-PCR and Western blot analysis. β-actin was used as the loading control. The blots are representative of three independent experiments with similar results. ( D ) Equal amounts of secreted protein from treated cells were incubated with a synthetic substrate labelled with amino-4-trifluoromethyl coumarin (AFC). The substrate is cleaved by cathepsin B to release AFC, which is fluorometrically detected. ( E ) The conditioned media from each sample was subjected to a chromogenic substrate, which is cleaved by active uPA and produces a colorimetrically detectable product. (F) The conditioned media was collected and separated on a non-reducing polyacrylamide gel containing gelatin A. Gelatinolytic activities are visualized as clear bands against the blue background of stained gelatin. The zymograms are representative of three independent experiments with similar results. The gels were cropped and the full-length gels are presented in Supplementary Fig. 3 . (G) The cells were placed into 8-μm porous culture inserts, treated with tivozanib and allowed to migrate for 48 h. The migrated cells on the lower surface of the inserts were fixed with methanol, stained with crystal violet, lysed with 30% acetic acid and the optical densitometry was measured at 590 nm. (H) For invasion assay, the cells were placed into matrigel-coated inserts and allowed to invade through the matrigel layer for 48 h. Data are given as mean ± SD. Statistically significant values of * p

    Article Snippet: Analysis of gene expression by quantitative reverse transcription-PCR The quantitative reverse transcription-PCR (qRT-PCR) analysis was performed on a LightCycler® 96 instrument (Roche Molecular Diagnostics) using RealQ SYBR Green PCR reagents (Ampliqon, Copenhagen, Denmark).

    Techniques: Staining, Quantitative RT-PCR, Western Blot, Incubation, Invasion Assay

    Confirmation of genes in SMEP by qRT-PCR. A heat map represents the expression levels for 29 selected genes using qRT-PCR from 151 SST-REX identified genes and 81 potential NSC SMEP by microarray. Each colored grid in the heat map represents the relative abundance of the transcript compared to the Gapdh level. Each gene was identified by either single or multiple niche cell type SMEP (parentheses on the left column). N: NSCs, T: TAPs, A: astrocytes, Ep: ependymal cells, C: choroid plexus, En: endothelial cells. * indicates the SMEP from microarray experiment.

    Journal: PLoS ONE

    Article Title: The Molecular Profiles of Neural Stem Cell Niche in the Adult Subventricular Zone

    doi: 10.1371/journal.pone.0050501

    Figure Lengend Snippet: Confirmation of genes in SMEP by qRT-PCR. A heat map represents the expression levels for 29 selected genes using qRT-PCR from 151 SST-REX identified genes and 81 potential NSC SMEP by microarray. Each colored grid in the heat map represents the relative abundance of the transcript compared to the Gapdh level. Each gene was identified by either single or multiple niche cell type SMEP (parentheses on the left column). N: NSCs, T: TAPs, A: astrocytes, Ep: ependymal cells, C: choroid plexus, En: endothelial cells. * indicates the SMEP from microarray experiment.

    Article Snippet: These preamplified products were diluted 200-fold and used for qRT-PCR analysis measuring SYBR Green incorporation (LightCycler 480 SYBR Green I Master, Roche) in a Roche Light Cycler 480 PCR machine.

    Techniques: Quantitative RT-PCR, Expressing, Microarray

    ER-α36 mediates postmenopausal low-level E 2 -induced inhibition of bone resorption and formation in cultured bone tissues. Normal or osteoporotic postmenopausal bone tissues were cultured in a medium containing 10% FBS and treated with vehicle, bone turnover–stimulating factors (50 ng/mL RANKL + 25 ng/mL M-CSF + 10 mM β-GP), or bone turnover–stimulating factors with various concentrations of E 2 for 28 days. mRNA levels of ER-α36, osteocalcin, and TRACP were determined using qRT-PCR and given as fold induction relative to vehicle-treated normal postmenopausal bone. The bars represent the mean ± SD. a p

    Journal: Journal of Bone and Mineral Research

    Article Title: Estrogen Receptor ?36 Mediates a Bone-Sparing Effect of 17?-Estrodiol in Postmenopausal Women

    doi: 10.1002/jbmr.169

    Figure Lengend Snippet: ER-α36 mediates postmenopausal low-level E 2 -induced inhibition of bone resorption and formation in cultured bone tissues. Normal or osteoporotic postmenopausal bone tissues were cultured in a medium containing 10% FBS and treated with vehicle, bone turnover–stimulating factors (50 ng/mL RANKL + 25 ng/mL M-CSF + 10 mM β-GP), or bone turnover–stimulating factors with various concentrations of E 2 for 28 days. mRNA levels of ER-α36, osteocalcin, and TRACP were determined using qRT-PCR and given as fold induction relative to vehicle-treated normal postmenopausal bone. The bars represent the mean ± SD. a p

    Article Snippet: Quantitative real-time PCR (qRT-PCR) analysis qRT-PCR was performed using a Roche Molecular Light Cycler (Roche Molecular Biochemicals).

    Techniques: Inhibition, Cell Culture, Quantitative RT-PCR

    The M2-K78R mutation affects influenza A virus replication. Viruses were used to infect A549 cells at an MOI of 1 for the following experiments. (A) A549 cells were incubated with influenza A viruses for 30 min at an MOI of 5. The cytoplasmic and nuclear viral RNAs were isolated and used for qRT-PCR. The percentage of vRNA was calculated as the ratio of the vRNA from each compartment to the vRNA from whole cells. Values represent the means ± SD of data from three independent experiments. (B) Infected A549 cells were harvested at the indicated times, and their vRNA levels of NP segments were determined by quantitative PCR. Values represent the means ± SD of data from three independent experiments. ***, P value of

    Journal: Journal of Virology

    Article Title: Ubiquitination of the Cytoplasmic Domain of Influenza A Virus M2 Protein Is Crucial for Production of Infectious Virus Particles

    doi: 10.1128/JVI.01972-17

    Figure Lengend Snippet: The M2-K78R mutation affects influenza A virus replication. Viruses were used to infect A549 cells at an MOI of 1 for the following experiments. (A) A549 cells were incubated with influenza A viruses for 30 min at an MOI of 5. The cytoplasmic and nuclear viral RNAs were isolated and used for qRT-PCR. The percentage of vRNA was calculated as the ratio of the vRNA from each compartment to the vRNA from whole cells. Values represent the means ± SD of data from three independent experiments. (B) Infected A549 cells were harvested at the indicated times, and their vRNA levels of NP segments were determined by quantitative PCR. Values represent the means ± SD of data from three independent experiments. ***, P value of

    Article Snippet: We followed the standard TaqMan method with the Universal Probe Library system (Roche) for quantitative RT-PCR (qRT-PCR) analysis.

    Techniques: Mutagenesis, Incubation, Isolation, Quantitative RT-PCR, Infection, Real-time Polymerase Chain Reaction

    The translation and transcription of flaB2 was restored in the Δ earA-sp and Δ 3A mutants. (A) Western blot analysis showed that FlaB2 was expressed in both Δ earA-sp and Δ 3A mutants, as well as in the wild-type cells but not in the Δ earA mutant or in a mutant deleted for flaB2 . (B) The transcription of flaB2 was restored in Δ earA-sp and Δ 3A strains, as detected by qRT-PCR experiments. While transcripts for flaB2 were barely detectable in the Δ earA mutant, flaB2 transcription in the Δ earA-sp and Δ 3A strains exceeded that of wild-type cells. Error bar shows standard derivation from nine data sets from three biological repeats, each of which were performed with triplicates.

    Journal: Frontiers in Microbiology

    Article Title: Bypassing the Need for the Transcriptional Activator EarA through a Spontaneous Deletion in the BRE Portion of the fla Operon Promoter in Methanococcus maripaludis

    doi: 10.3389/fmicb.2017.01329

    Figure Lengend Snippet: The translation and transcription of flaB2 was restored in the Δ earA-sp and Δ 3A mutants. (A) Western blot analysis showed that FlaB2 was expressed in both Δ earA-sp and Δ 3A mutants, as well as in the wild-type cells but not in the Δ earA mutant or in a mutant deleted for flaB2 . (B) The transcription of flaB2 was restored in Δ earA-sp and Δ 3A strains, as detected by qRT-PCR experiments. While transcripts for flaB2 were barely detectable in the Δ earA mutant, flaB2 transcription in the Δ earA-sp and Δ 3A strains exceeded that of wild-type cells. Error bar shows standard derivation from nine data sets from three biological repeats, each of which were performed with triplicates.

    Article Snippet: Quantitative RT-PCR (qRT-PCR) Analysis of the flaB2 Transcription Level in Mc. maripaludis Strains Total RNA from an Mc. maripaludis overnight cell culture was extracted using a High Pure RNA Isolation Kit (Roche Life Science) following a modified Gram negative bacteria RNA extraction protocol with an additional DNase treatment using a TURBO DNA-free Kit (Ambion) at 37°C for 30 min.

    Techniques: Western Blot, Mutagenesis, Quantitative RT-PCR

    Knockdown (KD) of Dachd in zebrafish larvae compromised the glomerular filtration barrier. A, Expression of Dachd and β‐actin in mesonephric glomeruli of the adult zebrafish imaged by laser scanning microscopy. Paraffin sections were stained with Hoechst ( DNA , blue) and with antibodies against β‐actin (green) and dachd (red). Scale bar represents 5 μm. B, Cryosections of 5 dpf wt1a:gfp zebrafish larvae showed a strong down‐regulation of Dachd expression after injection of dachd vivo morpholinos ( MO ). Scale bar represents 20 μm. C, Zebrafish eggs were injected with either control MO (Ctrl MO ), dachd splice or trans MO . Brightfield images of zebrafish larvae (3 dpf) showed oedema formation after dachd splice or trans MO injection (arrows in C; n = 3). Scale bars represent 250 μm. D, The dachd mRNA was determined by qRT ‐ PCR analysis. Total mRNA extracts of whole larvae (3 dpf) injected with Ctrl MO (white bar), dachd splice (grey bar) or trans MO (black bar) (mean ± SEM , n = 5, 25 larvae per group) were used for cDNA synthesis. Data sets were normalized to Ctrl MO ‐injected samples and the reference gene eef1a1l1 . ** P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: The transcription factor Dach1 is essential for podocyte function, et al. The transcription factor Dach1 is essential for podocyte function

    doi: 10.1111/jcmm.13544

    Figure Lengend Snippet: Knockdown (KD) of Dachd in zebrafish larvae compromised the glomerular filtration barrier. A, Expression of Dachd and β‐actin in mesonephric glomeruli of the adult zebrafish imaged by laser scanning microscopy. Paraffin sections were stained with Hoechst ( DNA , blue) and with antibodies against β‐actin (green) and dachd (red). Scale bar represents 5 μm. B, Cryosections of 5 dpf wt1a:gfp zebrafish larvae showed a strong down‐regulation of Dachd expression after injection of dachd vivo morpholinos ( MO ). Scale bar represents 20 μm. C, Zebrafish eggs were injected with either control MO (Ctrl MO ), dachd splice or trans MO . Brightfield images of zebrafish larvae (3 dpf) showed oedema formation after dachd splice or trans MO injection (arrows in C; n = 3). Scale bars represent 250 μm. D, The dachd mRNA was determined by qRT ‐ PCR analysis. Total mRNA extracts of whole larvae (3 dpf) injected with Ctrl MO (white bar), dachd splice (grey bar) or trans MO (black bar) (mean ± SEM , n = 5, 25 larvae per group) were used for cDNA synthesis. Data sets were normalized to Ctrl MO ‐injected samples and the reference gene eef1a1l1 . ** P

    Article Snippet: The quantitative real‐time PCR (qRT‐PCR) analysis was performed on a LightCycler Nano (Roche Diagnostics GmbH, Mannheim, Germany) using the iTaq Universal SYBR Green Supermix (Bio‐Rad, Hercules, CA, USA) with Actb and Gapdh as reference genes.

    Techniques: Filtration, Expressing, Laser-Scanning Microscopy, Staining, Injection, Quantitative RT-PCR

    Knockdown (KD) of Dachd in zebrafish larvae affected podocyte gene expression and induced foot process effacement. A, Cryosections of 3 dpf zebrafish larvae showed an increase in Bowman's space (arrows) after the KD of Dachd (n = 3). Nephrin staining is shown in the middle panel. Scale bar represents 20 μm. B, Electron microscopy of Dachd KD and Ctrl morpholinos ( MO ) zebrafish larvae. Ctrl MO larvae showed a normally structured filtration barrier with intact foot processes (arrows). Foot processes of dachd vivo MO larvae were disordered with microvilli‐like structures (arrowheads) protruding in an increased Bowman's space (Bo). glomerular basement membrane and endothelial cells were also affected by MO treatment (circle). Scale bars represent 10 μm in the upper and 1 μm in the lower pictures. P, podocyte; E, endothelial cell; Ca, capillary. C, The expression of podocyte genes ( synpo , nphs1 and nphs2 ) was determined by qRT ‐ PCR analysis. Total mRNA extracts of whole larvae (3 dpf) injected with Ctrl MO (white bars, means ± SD , n = 5), dachd splice MO (gray bars, means ± SD , n = 5) or dachd trans MO (black bars, means ± SD , n = 5) were used for cDNA synthesis. Data sets were normalized to Ctrl MO ‐injected samples and the reference genes rpl13 , eef1a1l1 and 18s RNA . **** P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: The transcription factor Dach1 is essential for podocyte function, et al. The transcription factor Dach1 is essential for podocyte function

    doi: 10.1111/jcmm.13544

    Figure Lengend Snippet: Knockdown (KD) of Dachd in zebrafish larvae affected podocyte gene expression and induced foot process effacement. A, Cryosections of 3 dpf zebrafish larvae showed an increase in Bowman's space (arrows) after the KD of Dachd (n = 3). Nephrin staining is shown in the middle panel. Scale bar represents 20 μm. B, Electron microscopy of Dachd KD and Ctrl morpholinos ( MO ) zebrafish larvae. Ctrl MO larvae showed a normally structured filtration barrier with intact foot processes (arrows). Foot processes of dachd vivo MO larvae were disordered with microvilli‐like structures (arrowheads) protruding in an increased Bowman's space (Bo). glomerular basement membrane and endothelial cells were also affected by MO treatment (circle). Scale bars represent 10 μm in the upper and 1 μm in the lower pictures. P, podocyte; E, endothelial cell; Ca, capillary. C, The expression of podocyte genes ( synpo , nphs1 and nphs2 ) was determined by qRT ‐ PCR analysis. Total mRNA extracts of whole larvae (3 dpf) injected with Ctrl MO (white bars, means ± SD , n = 5), dachd splice MO (gray bars, means ± SD , n = 5) or dachd trans MO (black bars, means ± SD , n = 5) were used for cDNA synthesis. Data sets were normalized to Ctrl MO ‐injected samples and the reference genes rpl13 , eef1a1l1 and 18s RNA . **** P

    Article Snippet: The quantitative real‐time PCR (qRT‐PCR) analysis was performed on a LightCycler Nano (Roche Diagnostics GmbH, Mannheim, Germany) using the iTaq Universal SYBR Green Supermix (Bio‐Rad, Hercules, CA, USA) with Actb and Gapdh as reference genes.

    Techniques: Expressing, Staining, Electron Microscopy, Filtration, Quantitative RT-PCR, Injection

    Dach1 and synaptopodin expression in mouse kidney and cell culture. A, Expression of Dach1 and synaptopodin in mouse glomeruli imaged by laser scanning microscopy (n = 3). Mouse kidney paraffin sections were stained with Hoechst ( DNA , blue) and with antibodies against synaptopodin (green) and Dach1 (red). Scale bar represents 25 μm. B, Dach1 and synaptopodin protein expression were compared between PEC s, podocytes (Podo), isolated glomeruli (Glom) and mouse kidney by Western blots (n = 3). Gapdh was used as a reference protein. C, Relative mRNA levels of Dach1 and Synpo in PEC s (white bars, means ± SD , n = 5) and podocytes (grey bars, means ± SD , n = 5) were determined by qRT ‐ PCR . Normalization was performed against PEC expression levels and Gapdh as reference gene. *** P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: The transcription factor Dach1 is essential for podocyte function, et al. The transcription factor Dach1 is essential for podocyte function

    doi: 10.1111/jcmm.13544

    Figure Lengend Snippet: Dach1 and synaptopodin expression in mouse kidney and cell culture. A, Expression of Dach1 and synaptopodin in mouse glomeruli imaged by laser scanning microscopy (n = 3). Mouse kidney paraffin sections were stained with Hoechst ( DNA , blue) and with antibodies against synaptopodin (green) and Dach1 (red). Scale bar represents 25 μm. B, Dach1 and synaptopodin protein expression were compared between PEC s, podocytes (Podo), isolated glomeruli (Glom) and mouse kidney by Western blots (n = 3). Gapdh was used as a reference protein. C, Relative mRNA levels of Dach1 and Synpo in PEC s (white bars, means ± SD , n = 5) and podocytes (grey bars, means ± SD , n = 5) were determined by qRT ‐ PCR . Normalization was performed against PEC expression levels and Gapdh as reference gene. *** P

    Article Snippet: The quantitative real‐time PCR (qRT‐PCR) analysis was performed on a LightCycler Nano (Roche Diagnostics GmbH, Mannheim, Germany) using the iTaq Universal SYBR Green Supermix (Bio‐Rad, Hercules, CA, USA) with Actb and Gapdh as reference genes.

    Techniques: Expressing, Cell Culture, Laser-Scanning Microscopy, Staining, Isolation, Western Blot, Quantitative RT-PCR

    Effect of Dach1 expression in PEC s on synaptopodin and further PEC /podocyte proteins. A, The up‐regulation of synaptopodin (red) and co‐localization with F‐actin (green) was shown by immunofluorescence and super resolution microscopy in PEC ‐Dach1. B, Synaptopodin‐positive cells are significantly increased in PEC ‐Dach1 as compared to PEC ‐Ctrl (upper diagram, n = 5 experiments with > 100 cells per experiment). Synaptopodin‐positive PEC s were almost always positive for Dach1 expression (lower diagram, n = 5 experiments with > 100 cells). C, Podocyte marker genes ( Wt1 , Nphs2 ), PEC marker genes ( Pax2 , Cav1 ) and differentiation/development regulators ( Eya1, Eya3 ) were quantified by qRT ‐ PCR analysis of total mRNA isolated from PEC ‐Ctrl and PEC ‐Dach1. qRT ‐ PCR experiments were normalized to PEC ‐Ctrl (dashed line). Actin , Gapdh and 18s RNA served as reference genes (n ≥ 3, means ± SEM ). D, WT 1 up‐regulation and Pax2 down‐regulation in PEC ‐Dach1 were shown by Western blot analysis. E, Relative Wt1 and Pax2 protein levels on Western blots were quantified for PEC ‐Ctrl (white bars) and PEC ‐Dach1 (black bars) and revealed an up‐regulation of WT 1 (n = 4, means ± SEM ) and a down‐regulation of Pax2 (n = 6, means ± SEM ) in PEC ‐Dach1. Western blots were normalized to PEC ‐Ctrl and Gapdh levels. *** P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: The transcription factor Dach1 is essential for podocyte function, et al. The transcription factor Dach1 is essential for podocyte function

    doi: 10.1111/jcmm.13544

    Figure Lengend Snippet: Effect of Dach1 expression in PEC s on synaptopodin and further PEC /podocyte proteins. A, The up‐regulation of synaptopodin (red) and co‐localization with F‐actin (green) was shown by immunofluorescence and super resolution microscopy in PEC ‐Dach1. B, Synaptopodin‐positive cells are significantly increased in PEC ‐Dach1 as compared to PEC ‐Ctrl (upper diagram, n = 5 experiments with > 100 cells per experiment). Synaptopodin‐positive PEC s were almost always positive for Dach1 expression (lower diagram, n = 5 experiments with > 100 cells). C, Podocyte marker genes ( Wt1 , Nphs2 ), PEC marker genes ( Pax2 , Cav1 ) and differentiation/development regulators ( Eya1, Eya3 ) were quantified by qRT ‐ PCR analysis of total mRNA isolated from PEC ‐Ctrl and PEC ‐Dach1. qRT ‐ PCR experiments were normalized to PEC ‐Ctrl (dashed line). Actin , Gapdh and 18s RNA served as reference genes (n ≥ 3, means ± SEM ). D, WT 1 up‐regulation and Pax2 down‐regulation in PEC ‐Dach1 were shown by Western blot analysis. E, Relative Wt1 and Pax2 protein levels on Western blots were quantified for PEC ‐Ctrl (white bars) and PEC ‐Dach1 (black bars) and revealed an up‐regulation of WT 1 (n = 4, means ± SEM ) and a down‐regulation of Pax2 (n = 6, means ± SEM ) in PEC ‐Dach1. Western blots were normalized to PEC ‐Ctrl and Gapdh levels. *** P

    Article Snippet: The quantitative real‐time PCR (qRT‐PCR) analysis was performed on a LightCycler Nano (Roche Diagnostics GmbH, Mannheim, Germany) using the iTaq Universal SYBR Green Supermix (Bio‐Rad, Hercules, CA, USA) with Actb and Gapdh as reference genes.

    Techniques: Expressing, Immunofluorescence, Microscopy, Marker, Quantitative RT-PCR, Isolation, Western Blot

    Expression analysis of three AhDGAT genes using qRT-PCR under different stresses. CL (0 h to 72 h), leaves exposed to cold (4°C) treatment. SL (0 h to 48 h), leaves exposed to high salt (200 mM NaCl) treatment. SR (0 h to 72 h), roots exposed to high salt (200 mM NaCl) treatment. DL (0 h to 72 h), leaves exposed to 20% PEG-6000 treatment. DR (0 h to 72 h), roots exposed to 20% PEG-6000 treatment. AL (0 h to 72 h), leaves exposed to 100 uM ABA treatment. AR (0 h to 72 h), roots exposed to 100 uM ABA treatment. The relative mRNA abundance was normalized with respect to the peanut AhACT11 gene. The bars were standard deviations (SD) of three technical repeats.

    Journal: PLoS ONE

    Article Title: Cloning and Functional Analysis of Three Diacylglycerol Acyltransferase Genes from Peanut (Arachis hypogaea L.)

    doi: 10.1371/journal.pone.0105834

    Figure Lengend Snippet: Expression analysis of three AhDGAT genes using qRT-PCR under different stresses. CL (0 h to 72 h), leaves exposed to cold (4°C) treatment. SL (0 h to 48 h), leaves exposed to high salt (200 mM NaCl) treatment. SR (0 h to 72 h), roots exposed to high salt (200 mM NaCl) treatment. DL (0 h to 72 h), leaves exposed to 20% PEG-6000 treatment. DR (0 h to 72 h), roots exposed to 20% PEG-6000 treatment. AL (0 h to 72 h), leaves exposed to 100 uM ABA treatment. AR (0 h to 72 h), roots exposed to 100 uM ABA treatment. The relative mRNA abundance was normalized with respect to the peanut AhACT11 gene. The bars were standard deviations (SD) of three technical repeats.

    Article Snippet: Quantitative real-time RT-PCR (qRT-PCR) qRT-PCR analysis was performed using a LightCycler 2.0 instrument system (Roche, Germany).

    Techniques: Expressing, Quantitative RT-PCR

    Expression analysis of three AhDGAT genes using qRT-PCR in seven peanut tissues and at six stages of seed development. DT (different tissues): R, root; SM, stem; L, leaf; C, cotyledons; H, hypocotyls; F, flower; SD, seed. DS (10 to 60 DAP): six developmental stages of seeds. The relative mRNA abundance was normalized with respect to the peanut AhACT11 gene. The bars were standard deviations (SD) of three technical repeats.

    Journal: PLoS ONE

    Article Title: Cloning and Functional Analysis of Three Diacylglycerol Acyltransferase Genes from Peanut (Arachis hypogaea L.)

    doi: 10.1371/journal.pone.0105834

    Figure Lengend Snippet: Expression analysis of three AhDGAT genes using qRT-PCR in seven peanut tissues and at six stages of seed development. DT (different tissues): R, root; SM, stem; L, leaf; C, cotyledons; H, hypocotyls; F, flower; SD, seed. DS (10 to 60 DAP): six developmental stages of seeds. The relative mRNA abundance was normalized with respect to the peanut AhACT11 gene. The bars were standard deviations (SD) of three technical repeats.

    Article Snippet: Quantitative real-time RT-PCR (qRT-PCR) qRT-PCR analysis was performed using a LightCycler 2.0 instrument system (Roche, Germany).

    Techniques: Expressing, Quantitative RT-PCR

    Pairwise variation (Vn/Vn+1) values in nine subsets calculated by using geNorm . The cut-off value to determine the optimal number of reference genes for qRT-PCR normalization is 0.15.

    Journal: Frontiers in Plant Science

    Article Title: Selection and Validation of Appropriate Reference Genes for Quantitative Real-Time PCR Analysis of Gene Expression in Lycoris aurea

    doi: 10.3389/fpls.2016.00536

    Figure Lengend Snippet: Pairwise variation (Vn/Vn+1) values in nine subsets calculated by using geNorm . The cut-off value to determine the optimal number of reference genes for qRT-PCR normalization is 0.15.

    Article Snippet: Quantitative real-time PCR analysis qRT-PCR was conducted in 96-well plates and performed on the LightCycler 480 (Roche Molecular Biochemicals, Mannheim, Germany).

    Techniques: Quantitative RT-PCR

    Validation of qRT-PCR results through comparison with RNA-seq expression profiles. (A) Stability ranking of candidate genes by CV of FPKM in RNA-seq. The gene with lower CV indicates more stable expression. (B) Correlation analysis between ranking of MeJA treatment subset by qRT-PCR and the ranking of RNA-seq. CV, coefficient of variation; FPKM, fragments per kilobase of exon model per million mapped reads.

    Journal: Frontiers in Plant Science

    Article Title: Selection and Validation of Appropriate Reference Genes for Quantitative Real-Time PCR Analysis of Gene Expression in Lycoris aurea

    doi: 10.3389/fpls.2016.00536

    Figure Lengend Snippet: Validation of qRT-PCR results through comparison with RNA-seq expression profiles. (A) Stability ranking of candidate genes by CV of FPKM in RNA-seq. The gene with lower CV indicates more stable expression. (B) Correlation analysis between ranking of MeJA treatment subset by qRT-PCR and the ranking of RNA-seq. CV, coefficient of variation; FPKM, fragments per kilobase of exon model per million mapped reads.

    Article Snippet: Quantitative real-time PCR analysis qRT-PCR was conducted in 96-well plates and performed on the LightCycler 480 (Roche Molecular Biochemicals, Mannheim, Germany).

    Techniques: Quantitative RT-PCR, RNA Sequencing Assay, Expressing

    Validation of the DEM (differentially expressed miRNAs) in autotetraploid and diploid rice at each pollen development stage (pre-meiotic interphase (PMA), meiosis (MA) and single microspore stage (SCP)). U6 snRNA was used as an internal reference for the qRT-PCR. The x - and y -axis represent the miRNAs and relative expression levels, respectively. Error bars represent the standard deviation (SD) of three biological replicates. “4x” and “2x” represent the autotetraploid and diploid rice, respectively.

    Journal: International Journal of Molecular Sciences

    Article Title: Comparative Small RNA Analysis of Pollen Development in Autotetraploid and Diploid Rice

    doi: 10.3390/ijms17040499

    Figure Lengend Snippet: Validation of the DEM (differentially expressed miRNAs) in autotetraploid and diploid rice at each pollen development stage (pre-meiotic interphase (PMA), meiosis (MA) and single microspore stage (SCP)). U6 snRNA was used as an internal reference for the qRT-PCR. The x - and y -axis represent the miRNAs and relative expression levels, respectively. Error bars represent the standard deviation (SD) of three biological replicates. “4x” and “2x” represent the autotetraploid and diploid rice, respectively.

    Article Snippet: Quantitative Real-Time PCR (qRT-PCR) Analysis The RNA was extracted from anthers of Taichung65-2x and Taichung65-4x at each stage of pollen development, and used as template for reverse transcription with miRNA-specific stem-loop RT primers [ ] using the Transcriptor First Strand cDNA Synthesis Kit (Roche, Mannheim, Germany).

    Techniques: Quantitative RT-PCR, Expressing, Standard Deviation

    Expression analysis of fatty acid desaturase genes of peanut using qRT-PCR in peanut leaves and roots under drought stress. 0h to 72h, leaves exposed to 20% PEG-6000 treatment. The relative mRNA abundance was normalized with respect to the peanut AhTUA5 gene. The bars were standard deviations (SD) of three biological repeats.

    Journal: PLoS ONE

    Article Title: Isolation and functional analysis of fatty acid desaturase genes from peanut (Arachis hypogaea L.)

    doi: 10.1371/journal.pone.0189759

    Figure Lengend Snippet: Expression analysis of fatty acid desaturase genes of peanut using qRT-PCR in peanut leaves and roots under drought stress. 0h to 72h, leaves exposed to 20% PEG-6000 treatment. The relative mRNA abundance was normalized with respect to the peanut AhTUA5 gene. The bars were standard deviations (SD) of three biological repeats.

    Article Snippet: Quantitative real-time RT-PCR qRT-PCR analysis was performed using a LightCycler 2.0 instrument system (Roche, Germany).

    Techniques: Expressing, Quantitative RT-PCR

    Expression analysis of fatty acid desaturase genes of peanut using qRT-PCR in peanut leaves upon cold treatment. 0h to 72h, leaves exposed to cold (4°C) treatment. The relative mRNA abundance was normalized with respect to the peanut AhTUA5 gene. The bars were standard deviations (SD) of three biological repeats.

    Journal: PLoS ONE

    Article Title: Isolation and functional analysis of fatty acid desaturase genes from peanut (Arachis hypogaea L.)

    doi: 10.1371/journal.pone.0189759

    Figure Lengend Snippet: Expression analysis of fatty acid desaturase genes of peanut using qRT-PCR in peanut leaves upon cold treatment. 0h to 72h, leaves exposed to cold (4°C) treatment. The relative mRNA abundance was normalized with respect to the peanut AhTUA5 gene. The bars were standard deviations (SD) of three biological repeats.

    Article Snippet: Quantitative real-time RT-PCR qRT-PCR analysis was performed using a LightCycler 2.0 instrument system (Roche, Germany).

    Techniques: Expressing, Quantitative RT-PCR

    Expression analysis of fatty acid desaturase genes of peanut using qRT-PCR in peanut leaves and roots under ABA treatment. 0h to 72h, leaves exposed to 100uM ABA treatment. The relative mRNA abundance was normalized with respect to the peanut AhTUA5 gene. The bars were standard deviations (SD) of three biological repeats.

    Journal: PLoS ONE

    Article Title: Isolation and functional analysis of fatty acid desaturase genes from peanut (Arachis hypogaea L.)

    doi: 10.1371/journal.pone.0189759

    Figure Lengend Snippet: Expression analysis of fatty acid desaturase genes of peanut using qRT-PCR in peanut leaves and roots under ABA treatment. 0h to 72h, leaves exposed to 100uM ABA treatment. The relative mRNA abundance was normalized with respect to the peanut AhTUA5 gene. The bars were standard deviations (SD) of three biological repeats.

    Article Snippet: Quantitative real-time RT-PCR qRT-PCR analysis was performed using a LightCycler 2.0 instrument system (Roche, Germany).

    Techniques: Expressing, Quantitative RT-PCR

    Expression analysis of fatty acid desaturase genes of peanut using qRT-PCR in peanut leaves and roots under salt stress. 0h to 48h, leaves exposed to high salt (200 mM NaCl) treatment. The relative mRNA abundance was normalized with respect to the peanut AhTUA5 gene. The bars were standard deviations (SD) of three biological repeats.

    Journal: PLoS ONE

    Article Title: Isolation and functional analysis of fatty acid desaturase genes from peanut (Arachis hypogaea L.)

    doi: 10.1371/journal.pone.0189759

    Figure Lengend Snippet: Expression analysis of fatty acid desaturase genes of peanut using qRT-PCR in peanut leaves and roots under salt stress. 0h to 48h, leaves exposed to high salt (200 mM NaCl) treatment. The relative mRNA abundance was normalized with respect to the peanut AhTUA5 gene. The bars were standard deviations (SD) of three biological repeats.

    Article Snippet: Quantitative real-time RT-PCR qRT-PCR analysis was performed using a LightCycler 2.0 instrument system (Roche, Germany).

    Techniques: Expressing, Quantitative RT-PCR

    Expression analysis of fatty acid desaturase genes of peanut using qRT-PCR at six stages of seed development. DS (10 to 60 DAP): six developmental stages of seeds. The relative mRNA abundance was normalized with respect to the peanut AhTUA5 gene. The bars were standard deviations (SD) of three biological repeats.

    Journal: PLoS ONE

    Article Title: Isolation and functional analysis of fatty acid desaturase genes from peanut (Arachis hypogaea L.)

    doi: 10.1371/journal.pone.0189759

    Figure Lengend Snippet: Expression analysis of fatty acid desaturase genes of peanut using qRT-PCR at six stages of seed development. DS (10 to 60 DAP): six developmental stages of seeds. The relative mRNA abundance was normalized with respect to the peanut AhTUA5 gene. The bars were standard deviations (SD) of three biological repeats.

    Article Snippet: Quantitative real-time RT-PCR qRT-PCR analysis was performed using a LightCycler 2.0 instrument system (Roche, Germany).

    Techniques: Expressing, Quantitative RT-PCR

    Expression analysis of fatty acid desaturase genes of peanut using qRT-PCR in five peanut tissues. R, root; SM, stem; L, leaf; F, flower; SD, seed. The relative mRNA abundance was normalized with respect to the peanut AhTUA5 gene. The bars were standard deviations (SD) of three biological repeats.

    Journal: PLoS ONE

    Article Title: Isolation and functional analysis of fatty acid desaturase genes from peanut (Arachis hypogaea L.)

    doi: 10.1371/journal.pone.0189759

    Figure Lengend Snippet: Expression analysis of fatty acid desaturase genes of peanut using qRT-PCR in five peanut tissues. R, root; SM, stem; L, leaf; F, flower; SD, seed. The relative mRNA abundance was normalized with respect to the peanut AhTUA5 gene. The bars were standard deviations (SD) of three biological repeats.

    Article Snippet: Quantitative real-time RT-PCR qRT-PCR analysis was performed using a LightCycler 2.0 instrument system (Roche, Germany).

    Techniques: Expressing, Quantitative RT-PCR

    Validation of ABA- and JA-regulated genes using qRT-PCR. The ΔΔCT value method was used to determine the relative fold changes. All data were normalized to the expression level of Ubi5 . Error bars represent standard error of three replicates.

    Journal: Current Genomics

    Article Title: Transcriptome Analysis of ABA/JA-Dual Responsive Genes in Rice Shoot and Root

    doi: 10.2174/1389202918666170228134205

    Figure Lengend Snippet: Validation of ABA- and JA-regulated genes using qRT-PCR. The ΔΔCT value method was used to determine the relative fold changes. All data were normalized to the expression level of Ubi5 . Error bars represent standard error of three replicates.

    Article Snippet: Real-Time Quantitative RT-PCR Analysis Real-time quantitative RT-PCR (qRT-PCR) was performed using LightCycler® 480 SYBR Green I Master (Roche, Swiss) and the MyiQ (Bio-Rad, US).

    Techniques: Quantitative RT-PCR, Expressing

    Validation of ABA regulation of genes expressed in root and shoot using qRT-PCR. The ΔΔCT method was used to determine the relative fold change. All data were normalized to the expression level of Ubi5 . Error bars represent standard error of three replicates.

    Journal: Current Genomics

    Article Title: Transcriptome Analysis of ABA/JA-Dual Responsive Genes in Rice Shoot and Root

    doi: 10.2174/1389202918666170228134205

    Figure Lengend Snippet: Validation of ABA regulation of genes expressed in root and shoot using qRT-PCR. The ΔΔCT method was used to determine the relative fold change. All data were normalized to the expression level of Ubi5 . Error bars represent standard error of three replicates.

    Article Snippet: Real-Time Quantitative RT-PCR Analysis Real-time quantitative RT-PCR (qRT-PCR) was performed using LightCycler® 480 SYBR Green I Master (Roche, Swiss) and the MyiQ (Bio-Rad, US).

    Techniques: Quantitative RT-PCR, Expressing

    Transcript levels of cbs are up-regulated by 1,25(OH) 2 D 3 in murine MC3T3-E1 and primary osteoblasts. (A) Transcript levels were determined by qRT-PCR at different time points after treatment with 1,25(OH) 2 D 3 (10 −8 M). Each data point represents

    Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

    Article Title: 1,25-dihydroxyvitamin D3 influences cellular homocysteine levels in murine pre-osteoblastic MC3T3-E1 cells by direct regulation of cystathionine ?-synthase

    doi: 10.1002/jbmr.493

    Figure Lengend Snippet: Transcript levels of cbs are up-regulated by 1,25(OH) 2 D 3 in murine MC3T3-E1 and primary osteoblasts. (A) Transcript levels were determined by qRT-PCR at different time points after treatment with 1,25(OH) 2 D 3 (10 −8 M). Each data point represents

    Article Snippet: Total RNA for quantitative real-time polymerase chain reaction (qRT-PCR) analysis was isolated with the High Pure RNA Isolation Kit (Roche) following the manufacturer’s instructions.

    Techniques: Quantitative RT-PCR

    Expression profiles of loquat sucrose synthase and sucrose phosphate synthase ( EjSPS and EjSS ) using qRT-PCR during fruit development. Each point is the mean of three determinations. Vertical bars, if larger than symbols, represent the standard error of the mean ( n = 3).

    Journal: International Journal of Molecular Sciences

    Article Title: Comparative Transcriptional Analysis of Loquat Fruit Identifies Major Signal Networks Involved in Fruit Development and Ripening Process

    doi: 10.3390/ijms17111837

    Figure Lengend Snippet: Expression profiles of loquat sucrose synthase and sucrose phosphate synthase ( EjSPS and EjSS ) using qRT-PCR during fruit development. Each point is the mean of three determinations. Vertical bars, if larger than symbols, represent the standard error of the mean ( n = 3).

    Article Snippet: Gene Expression Analysis by qRT-PCR qRT-PCR was performed using the first-strand cDNA as templates on a LightCycler® 480 Real-Time PCR system (Roche Diagnostics, Vienna, Austria), with 10 µL FastStart Universal SYBR Green I Master (ROX, Roche Diagnostics, Mannheim, Germany), 1 µL forward primer (10 µmol·L−1 ), 1 µL reverse primer (10 µmol·L−1 ), 1 µL cDNA (10 ng) and 7 µL water.

    Techniques: Expressing, Quantitative RT-PCR

    Expression profiles of differentially expressed genes DEGs involved in cell wall loosening and enlargement using qRT-PCR. Each point is the mean of three determinations. Vertical bars, if larger than symbols, represent the standard error of the mean ( n = 3).

    Journal: International Journal of Molecular Sciences

    Article Title: Comparative Transcriptional Analysis of Loquat Fruit Identifies Major Signal Networks Involved in Fruit Development and Ripening Process

    doi: 10.3390/ijms17111837

    Figure Lengend Snippet: Expression profiles of differentially expressed genes DEGs involved in cell wall loosening and enlargement using qRT-PCR. Each point is the mean of three determinations. Vertical bars, if larger than symbols, represent the standard error of the mean ( n = 3).

    Article Snippet: Gene Expression Analysis by qRT-PCR qRT-PCR was performed using the first-strand cDNA as templates on a LightCycler® 480 Real-Time PCR system (Roche Diagnostics, Vienna, Austria), with 10 µL FastStart Universal SYBR Green I Master (ROX, Roche Diagnostics, Mannheim, Germany), 1 µL forward primer (10 µmol·L−1 ), 1 µL reverse primer (10 µmol·L−1 ), 1 µL cDNA (10 ng) and 7 µL water.

    Techniques: Expressing, Quantitative RT-PCR

    Distributions and expression profiles of transcription factors (TFs) during loquat fruit development. ( A ) Distributions and types of TFs at stage I; ( B ) Distributions and types of TFs at stage II; ( C ) Distributions and types of TFs at stage III; ( D ) Distributions and types of the differentially expressed TFs at three stages; ( E ) Expression profiles of the randomly selected TFs investigated by qRT-PCR. Each point is the mean of three determinations. Vertical bars represent the standard error of the mean ( n = 3). bHLH, basic helix-loop-helix; bZIP, leucine zipper, ZF, zinc finger; HSF, hot shock factor; NAC, cup-shaped cotyledon; AP2/ERF, APETALA2/ethylene responsive factor; AUX/IAA, Auxin/indole-3-acetic acid; EIN3/EIL, ethylene-insensitive 3 and EIN3-like.

    Journal: International Journal of Molecular Sciences

    Article Title: Comparative Transcriptional Analysis of Loquat Fruit Identifies Major Signal Networks Involved in Fruit Development and Ripening Process

    doi: 10.3390/ijms17111837

    Figure Lengend Snippet: Distributions and expression profiles of transcription factors (TFs) during loquat fruit development. ( A ) Distributions and types of TFs at stage I; ( B ) Distributions and types of TFs at stage II; ( C ) Distributions and types of TFs at stage III; ( D ) Distributions and types of the differentially expressed TFs at three stages; ( E ) Expression profiles of the randomly selected TFs investigated by qRT-PCR. Each point is the mean of three determinations. Vertical bars represent the standard error of the mean ( n = 3). bHLH, basic helix-loop-helix; bZIP, leucine zipper, ZF, zinc finger; HSF, hot shock factor; NAC, cup-shaped cotyledon; AP2/ERF, APETALA2/ethylene responsive factor; AUX/IAA, Auxin/indole-3-acetic acid; EIN3/EIL, ethylene-insensitive 3 and EIN3-like.

    Article Snippet: Gene Expression Analysis by qRT-PCR qRT-PCR was performed using the first-strand cDNA as templates on a LightCycler® 480 Real-Time PCR system (Roche Diagnostics, Vienna, Austria), with 10 µL FastStart Universal SYBR Green I Master (ROX, Roche Diagnostics, Mannheim, Germany), 1 µL forward primer (10 µmol·L−1 ), 1 µL reverse primer (10 µmol·L−1 ), 1 µL cDNA (10 ng) and 7 µL water.

    Techniques: Expressing, Quantitative RT-PCR

    Transcription profiles of differentially expressed genes DEGs involved in the ethylene signal pathway using qRT-PCR. Each point is the mean of three determinations. Vertical bars, if larger than symbols, represent the standard error of the mean ( n = 3).

    Journal: International Journal of Molecular Sciences

    Article Title: Comparative Transcriptional Analysis of Loquat Fruit Identifies Major Signal Networks Involved in Fruit Development and Ripening Process

    doi: 10.3390/ijms17111837

    Figure Lengend Snippet: Transcription profiles of differentially expressed genes DEGs involved in the ethylene signal pathway using qRT-PCR. Each point is the mean of three determinations. Vertical bars, if larger than symbols, represent the standard error of the mean ( n = 3).

    Article Snippet: Gene Expression Analysis by qRT-PCR qRT-PCR was performed using the first-strand cDNA as templates on a LightCycler® 480 Real-Time PCR system (Roche Diagnostics, Vienna, Austria), with 10 µL FastStart Universal SYBR Green I Master (ROX, Roche Diagnostics, Mannheim, Germany), 1 µL forward primer (10 µmol·L−1 ), 1 µL reverse primer (10 µmol·L−1 ), 1 µL cDNA (10 ng) and 7 µL water.

    Techniques: Quantitative RT-PCR

    Expression profiles of differentially expressed genes DEGs involved in the auxin signal pathway using qRT-PCR. Each point is the mean of three determinations. Vertical bars, if larger than symbols, represent the standard error of the mean ( n = 3).

    Journal: International Journal of Molecular Sciences

    Article Title: Comparative Transcriptional Analysis of Loquat Fruit Identifies Major Signal Networks Involved in Fruit Development and Ripening Process

    doi: 10.3390/ijms17111837

    Figure Lengend Snippet: Expression profiles of differentially expressed genes DEGs involved in the auxin signal pathway using qRT-PCR. Each point is the mean of three determinations. Vertical bars, if larger than symbols, represent the standard error of the mean ( n = 3).

    Article Snippet: Gene Expression Analysis by qRT-PCR qRT-PCR was performed using the first-strand cDNA as templates on a LightCycler® 480 Real-Time PCR system (Roche Diagnostics, Vienna, Austria), with 10 µL FastStart Universal SYBR Green I Master (ROX, Roche Diagnostics, Mannheim, Germany), 1 µL forward primer (10 µmol·L−1 ), 1 µL reverse primer (10 µmol·L−1 ), 1 µL cDNA (10 ng) and 7 µL water.

    Techniques: Expressing, Quantitative RT-PCR

    Comparison between DT and DS during drought stress and after rewatering. ( a ) DEGs in DT versus DS at various drought stress time points. ( b ) A venn diagram depicting the common and unique DEGs between d 1 and d 3 in DT and DS. ( c ) Expression patterns of the 61 common DEGs between DT and DS under drought conditions. ( d ) Correlation between transcriptome data and qRT-PCR results based on log 2 fold change of 10 selected genes.

    Journal: Scientific Reports

    Article Title: Transcriptomic, biochemical and physio-anatomical investigations shed more light on responses to drought stress in two contrasting sesame genotypes

    doi: 10.1038/s41598-017-09397-6

    Figure Lengend Snippet: Comparison between DT and DS during drought stress and after rewatering. ( a ) DEGs in DT versus DS at various drought stress time points. ( b ) A venn diagram depicting the common and unique DEGs between d 1 and d 3 in DT and DS. ( c ) Expression patterns of the 61 common DEGs between DT and DS under drought conditions. ( d ) Correlation between transcriptome data and qRT-PCR results based on log 2 fold change of 10 selected genes.

    Article Snippet: Validation of gene expression using qRT-PCR qRT-PCR analyses of target genes were performed according to Dossa et al . using a Light Cycler 480 II (Roche, Switzerland).

    Techniques: Expressing, Quantitative RT-PCR

    Opg-independent increase of osteclastogenesis in Ctnnb1 fl/fl /LysM +/cre mice. (A) TRAP activity staining on spine sections shows increased osteoclastogenesis in Ctnnb1 fl/fl /LysM +/cre mice. Histomorphometric quantification of osteoclast number (Oc.N/B.Pm) and osteoclast surface (OcS/BS) is given on the right. Bars, 50 µm. (B) Concentrations of collagen degradation products (Crosslaps), Opg, and Rankl in the serum of Ctnnb1 fl/fl /LysM +/+ and Ctnnb1 fl/fl /LysM +/cre mice. All error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences between the two groups. (C) qRT-PCR expression analysis of the indicated genes in marrow-flushed bones (left) or in bone marrow–derived osteoclast cultures (right) from Ctnnb1 fl/fl /LysM +/cre mice relative to Ctnnb1 fl/fl /LysM +/+ littermates. Error bars represent mean ± SD ( n = 4). (D) Quantification of TRAP-positive multinucleated cells in Ctnnb1 fl/fl /LysM +/+ and Ctnnb1 fl/fl /LysM +/cre bone marrow cells differentiated by addition of M-Csf, Rankl in the absence of presence of Wnt3a, as indicated. All error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences compared with untreated controls.

    Journal: The Journal of Cell Biology

    Article Title: Canonical Wnt signaling inhibits osteoclastogenesis independent of osteoprotegerin

    doi: 10.1083/jcb.201207142

    Figure Lengend Snippet: Opg-independent increase of osteclastogenesis in Ctnnb1 fl/fl /LysM +/cre mice. (A) TRAP activity staining on spine sections shows increased osteoclastogenesis in Ctnnb1 fl/fl /LysM +/cre mice. Histomorphometric quantification of osteoclast number (Oc.N/B.Pm) and osteoclast surface (OcS/BS) is given on the right. Bars, 50 µm. (B) Concentrations of collagen degradation products (Crosslaps), Opg, and Rankl in the serum of Ctnnb1 fl/fl /LysM +/+ and Ctnnb1 fl/fl /LysM +/cre mice. All error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences between the two groups. (C) qRT-PCR expression analysis of the indicated genes in marrow-flushed bones (left) or in bone marrow–derived osteoclast cultures (right) from Ctnnb1 fl/fl /LysM +/cre mice relative to Ctnnb1 fl/fl /LysM +/+ littermates. Error bars represent mean ± SD ( n = 4). (D) Quantification of TRAP-positive multinucleated cells in Ctnnb1 fl/fl /LysM +/+ and Ctnnb1 fl/fl /LysM +/cre bone marrow cells differentiated by addition of M-Csf, Rankl in the absence of presence of Wnt3a, as indicated. All error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences compared with untreated controls.

    Article Snippet: Whereas RNA was isolated after 6 h for qRT-PCR expression analysis, protein extracts were isolated after 30 min by cell lysis with RIPA buffer (1% NP-40, 1% sodium deoxycholate, 0.1% sodium dodecylsulfate, 150 mM sodium chloride, 2 mM EDTA, and 10 mM sodium phosphate) containing a protease and phosphatase inhibitor cocktail (Roche).

    Techniques: Mouse Assay, Activity Assay, Staining, Quantitative RT-PCR, Expressing, Derivative Assay

    Inhibition of osteoclastogenesis by canonical Wnt signaling. (A) Quantification of TRAP-positive multinucleated cells generated in the presence of different concentrations of LiCl, Wnt3a, or Wnt5a as indicated. (B) Quantification of TRAP-positive multinucleated cells generated after 7 d of culture in the permanent presence of recombinant Wnt3a and Rankl as indicated. (C) Quantification of TRAP-positive multinucleated cells generated after 7 d of culture, when Wnt3a (or LiCl) and Rankl were added from day 4 of culture. (D) qRT-PCR expression analysis for the indicated genes in bone marrow cultures at day 3 of differentiation after treatment with 200 ng/ml Wnt3a for 6 h. All error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences compared with controls. (E) Western blot analysis for phosphorylated Lrp6 and β-catenin after stimulation of bone marrow cultures at day 3 of differentiation by Wnt3a in the absence or presence of the Fzd8-CRD. The black arrows indicate the position and molecular weight of the nearest marker.

    Journal: The Journal of Cell Biology

    Article Title: Canonical Wnt signaling inhibits osteoclastogenesis independent of osteoprotegerin

    doi: 10.1083/jcb.201207142

    Figure Lengend Snippet: Inhibition of osteoclastogenesis by canonical Wnt signaling. (A) Quantification of TRAP-positive multinucleated cells generated in the presence of different concentrations of LiCl, Wnt3a, or Wnt5a as indicated. (B) Quantification of TRAP-positive multinucleated cells generated after 7 d of culture in the permanent presence of recombinant Wnt3a and Rankl as indicated. (C) Quantification of TRAP-positive multinucleated cells generated after 7 d of culture, when Wnt3a (or LiCl) and Rankl were added from day 4 of culture. (D) qRT-PCR expression analysis for the indicated genes in bone marrow cultures at day 3 of differentiation after treatment with 200 ng/ml Wnt3a for 6 h. All error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences compared with controls. (E) Western blot analysis for phosphorylated Lrp6 and β-catenin after stimulation of bone marrow cultures at day 3 of differentiation by Wnt3a in the absence or presence of the Fzd8-CRD. The black arrows indicate the position and molecular weight of the nearest marker.

    Article Snippet: Whereas RNA was isolated after 6 h for qRT-PCR expression analysis, protein extracts were isolated after 30 min by cell lysis with RIPA buffer (1% NP-40, 1% sodium deoxycholate, 0.1% sodium dodecylsulfate, 150 mM sodium chloride, 2 mM EDTA, and 10 mM sodium phosphate) containing a protease and phosphatase inhibitor cocktail (Roche).

    Techniques: Inhibition, Generated, Recombinant, Quantitative RT-PCR, Expressing, Western Blot, Molecular Weight, Marker

    Increased osteclastogenesis in Fzd8 -deficient mice. (A) Quantification of osteoclast number per bone perimeter (Oc.N/B.Pm) and osteoclast surface per bone surface (OcS/BS) in 24-wk-old wild-type and Fzd8 -deficient mice. Error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences between the two groups. (B) Western blot analysis for phosphorylated Lrp6 and β-catenin after stimulation of wild-type and Fzd8 -deficient osteoblasts by Wnt3a at day 10 of differentiation. The black arrows indicate the position and molecular weight of the nearest marker. (C) qRT-PCR expression analysis of the indicated genes in Fzd8 -deficient osteoblasts relative to wild-type cultures. Error bars represent mean ± SD ( n = 4). (D) qRT-PCR expression analysis of the same genes in Fzd8 -deficient marrow-flushed bones relative to wild-type littermates. Error bars represent mean ± SD ( n = 4). (E) Serum levels of Opg and Rankl in 24-wk-old wild-type and Fzd8 -deficient littermates. Error bars represent mean ± SD ( n = 6). (F) TRAP activity staining of bone marrow cells differentiated in the presence of M-Csf and Rankl with or without Wnt3a as indicated. Bars, 50 µm. Quantification of TRAP-positive multinucleated cells in wild-type and Fzd8 -deficient cultures is given on the right. Error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences compared with control-treated cells.

    Journal: The Journal of Cell Biology

    Article Title: Canonical Wnt signaling inhibits osteoclastogenesis independent of osteoprotegerin

    doi: 10.1083/jcb.201207142

    Figure Lengend Snippet: Increased osteclastogenesis in Fzd8 -deficient mice. (A) Quantification of osteoclast number per bone perimeter (Oc.N/B.Pm) and osteoclast surface per bone surface (OcS/BS) in 24-wk-old wild-type and Fzd8 -deficient mice. Error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences between the two groups. (B) Western blot analysis for phosphorylated Lrp6 and β-catenin after stimulation of wild-type and Fzd8 -deficient osteoblasts by Wnt3a at day 10 of differentiation. The black arrows indicate the position and molecular weight of the nearest marker. (C) qRT-PCR expression analysis of the indicated genes in Fzd8 -deficient osteoblasts relative to wild-type cultures. Error bars represent mean ± SD ( n = 4). (D) qRT-PCR expression analysis of the same genes in Fzd8 -deficient marrow-flushed bones relative to wild-type littermates. Error bars represent mean ± SD ( n = 4). (E) Serum levels of Opg and Rankl in 24-wk-old wild-type and Fzd8 -deficient littermates. Error bars represent mean ± SD ( n = 6). (F) TRAP activity staining of bone marrow cells differentiated in the presence of M-Csf and Rankl with or without Wnt3a as indicated. Bars, 50 µm. Quantification of TRAP-positive multinucleated cells in wild-type and Fzd8 -deficient cultures is given on the right. Error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences compared with control-treated cells.

    Article Snippet: Whereas RNA was isolated after 6 h for qRT-PCR expression analysis, protein extracts were isolated after 30 min by cell lysis with RIPA buffer (1% NP-40, 1% sodium deoxycholate, 0.1% sodium dodecylsulfate, 150 mM sodium chloride, 2 mM EDTA, and 10 mM sodium phosphate) containing a protease and phosphatase inhibitor cocktail (Roche).

    Techniques: Mouse Assay, Western Blot, Molecular Weight, Marker, Quantitative RT-PCR, Expressing, Activity Assay, Staining

    Compensatory induction of Fzd genes in Wnt3a-treated Fzd8 -deficient osteoclast cultures. (A) qRT-PCR expression analysis of the indicated genes in wild-type and Fzd8 -deficient bone marrow cultures differentiated in the presence of M-Csf and Rankl with or without Wnt3a as indicated. Error bars represent mean ± SD ( n = 4). Asterisks indicate statistically significant differences compared with control-treated cells. Number signs indicate statistically significant differences between wild-type and Fzd8 -deficient cultures. (B) qRT-PCR expression analysis of the indicated genes in Fzd8 -deficient marrow-flushed bones relative to wild-type littermates. Error bars represent mean ± SD ( n = 4).

    Journal: The Journal of Cell Biology

    Article Title: Canonical Wnt signaling inhibits osteoclastogenesis independent of osteoprotegerin

    doi: 10.1083/jcb.201207142

    Figure Lengend Snippet: Compensatory induction of Fzd genes in Wnt3a-treated Fzd8 -deficient osteoclast cultures. (A) qRT-PCR expression analysis of the indicated genes in wild-type and Fzd8 -deficient bone marrow cultures differentiated in the presence of M-Csf and Rankl with or without Wnt3a as indicated. Error bars represent mean ± SD ( n = 4). Asterisks indicate statistically significant differences compared with control-treated cells. Number signs indicate statistically significant differences between wild-type and Fzd8 -deficient cultures. (B) qRT-PCR expression analysis of the indicated genes in Fzd8 -deficient marrow-flushed bones relative to wild-type littermates. Error bars represent mean ± SD ( n = 4).

    Article Snippet: Whereas RNA was isolated after 6 h for qRT-PCR expression analysis, protein extracts were isolated after 30 min by cell lysis with RIPA buffer (1% NP-40, 1% sodium deoxycholate, 0.1% sodium dodecylsulfate, 150 mM sodium chloride, 2 mM EDTA, and 10 mM sodium phosphate) containing a protease and phosphatase inhibitor cocktail (Roche).

    Techniques: Quantitative RT-PCR, Expressing

    Fzd8 expression in bone cells. (A) qRT-PCR expression analysis for Wnt receptor-encoding genes using cDNA from various tissues and bone cells. Expression levels were normalized to Gapdh as indicated. Error bars represent mean ± SD ( n = 4). (B) qRT-PCR expression analysis for Fzd8 and Fzd9 during the course of osteoclast and osteoblast differentiation. Error bars represent mean ± SD ( n = 4). Asterisks indicate statistically significant differences compared with the earliest stage of differentiation. (C) Immunohistochemistry on human bone sections demonstrates the presence of FZD8 in osteoblasts (top) and osteoclasts (bottom). Bars, 25 µm.

    Journal: The Journal of Cell Biology

    Article Title: Canonical Wnt signaling inhibits osteoclastogenesis independent of osteoprotegerin

    doi: 10.1083/jcb.201207142

    Figure Lengend Snippet: Fzd8 expression in bone cells. (A) qRT-PCR expression analysis for Wnt receptor-encoding genes using cDNA from various tissues and bone cells. Expression levels were normalized to Gapdh as indicated. Error bars represent mean ± SD ( n = 4). (B) qRT-PCR expression analysis for Fzd8 and Fzd9 during the course of osteoclast and osteoblast differentiation. Error bars represent mean ± SD ( n = 4). Asterisks indicate statistically significant differences compared with the earliest stage of differentiation. (C) Immunohistochemistry on human bone sections demonstrates the presence of FZD8 in osteoblasts (top) and osteoclasts (bottom). Bars, 25 µm.

    Article Snippet: Whereas RNA was isolated after 6 h for qRT-PCR expression analysis, protein extracts were isolated after 30 min by cell lysis with RIPA buffer (1% NP-40, 1% sodium deoxycholate, 0.1% sodium dodecylsulfate, 150 mM sodium chloride, 2 mM EDTA, and 10 mM sodium phosphate) containing a protease and phosphatase inhibitor cocktail (Roche).

    Techniques: Expressing, Quantitative RT-PCR, Immunohistochemistry

    Opg-independent inhibition of osteoclastogenesis by Wnt3a. (A) Quantification of TRAP-positive multinucleated cells generated after 7 d of culture in the presence of Wnt3a and/or an anti-Opg antibody. Error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences compared with controls. (B) Quantification of TRAP-positive multinucleated cells generated from unsorted or CD11b-purified bone marrow cells after 7 d of culture in the presence of Wnt3a. Error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences compared with controls. (C) qRT-PCR expression analysis of the indicated genes in CD11b-purified bone marrow cells after treatment with 200 ng/ml Wnt3a for 6 h relative to untreated cells. Error bars represent mean ± SD ( n = 4). Asterisks indicate statistically significant differences compared with controls.

    Journal: The Journal of Cell Biology

    Article Title: Canonical Wnt signaling inhibits osteoclastogenesis independent of osteoprotegerin

    doi: 10.1083/jcb.201207142

    Figure Lengend Snippet: Opg-independent inhibition of osteoclastogenesis by Wnt3a. (A) Quantification of TRAP-positive multinucleated cells generated after 7 d of culture in the presence of Wnt3a and/or an anti-Opg antibody. Error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences compared with controls. (B) Quantification of TRAP-positive multinucleated cells generated from unsorted or CD11b-purified bone marrow cells after 7 d of culture in the presence of Wnt3a. Error bars represent mean ± SD ( n = 6). Asterisks indicate statistically significant differences compared with controls. (C) qRT-PCR expression analysis of the indicated genes in CD11b-purified bone marrow cells after treatment with 200 ng/ml Wnt3a for 6 h relative to untreated cells. Error bars represent mean ± SD ( n = 4). Asterisks indicate statistically significant differences compared with controls.

    Article Snippet: Whereas RNA was isolated after 6 h for qRT-PCR expression analysis, protein extracts were isolated after 30 min by cell lysis with RIPA buffer (1% NP-40, 1% sodium deoxycholate, 0.1% sodium dodecylsulfate, 150 mM sodium chloride, 2 mM EDTA, and 10 mM sodium phosphate) containing a protease and phosphatase inhibitor cocktail (Roche).

    Techniques: Inhibition, Generated, Purification, Quantitative RT-PCR, Expressing

    Validation of differentially expressed miRNAs and some of their targets during the seed germination in sacred lotus by qRT-PCR a Nnu-miR157a, b Nnu-miR168a-5p, c Nnu-miR393A-5p, d Nnu-miR396b-5p, e Nnu-miR397, f Nnu-miR2111a, g Nnu-miR156a, h Nnu-miR319c, i Nnu-miR394a, j Target of Nnu-miR156a (NNU_02036-RA, SPL16), k Target of Nnu-miR319c (NNU_00026-RA, TCP2), l Target of Nnu-miR394a (NNU_22183-RA, FBX)

    Journal: BMC Genomics

    Article Title: Small RNA and degradome profiling reveals miRNA regulation in the seed germination of ancient eudicot Nelumbo nucifera

    doi: 10.1186/s12864-016-3032-4

    Figure Lengend Snippet: Validation of differentially expressed miRNAs and some of their targets during the seed germination in sacred lotus by qRT-PCR a Nnu-miR157a, b Nnu-miR168a-5p, c Nnu-miR393A-5p, d Nnu-miR396b-5p, e Nnu-miR397, f Nnu-miR2111a, g Nnu-miR156a, h Nnu-miR319c, i Nnu-miR394a, j Target of Nnu-miR156a (NNU_02036-RA, SPL16), k Target of Nnu-miR319c (NNU_00026-RA, TCP2), l Target of Nnu-miR394a (NNU_22183-RA, FBX)

    Article Snippet: Real time qRT-PCR analysis of the miRNA and their targets was performed using the FastStart Universal SYBR Green Master Mix (Roche) on the StepOne plus PCR platform (Applied Biosystems).

    Techniques: Quantitative RT-PCR

    MED29 expression leads to reduced growth of NIH/3T3 cells in vitro . NIH/3T3 mouse fibroblast cell line and Hs 700T and MIA PaCa-2 human pancreatic cancer cell lines were stably transfected with MED29 or an empty control vector. ( a ) Relative MED29 mRNA expression levels were assessed by qRT-PCR in MED29 transduced Hs 700T and MIA PaCa-2 cells and their respective empty vector (mock) control cells. MED29 expression values were normalized against a TATA-box binding protein house-keeping gene. ( b ) Western blot was used to detect MED29 protein (21 kDa) in stable MED29-expressing cells vs. mock control cells. β-Tubulin was used as a loading control. ( c ) The growth of the NIH3T3/MED29-1 and NIH3T3/mock cells was monitored at indicated time points in normal growth medium and ( d ) after 24h of serum starvation (1% FBS). ( e ) The growth of the MIAPaCa2/MED29 cells were monitored at indicated time points after release from G1-arrest. The mean +/− s.d. of six replicates are shown. The experiments were repeated three times with similar results. * P

    Journal: International journal of cancer. Journal international du cancer

    Article Title: MED29, a component of the Mediator complex, possesses both oncogenic and tumor suppressive characteristics in pancreatic cancer

    doi: 10.1002/ijc.25924

    Figure Lengend Snippet: MED29 expression leads to reduced growth of NIH/3T3 cells in vitro . NIH/3T3 mouse fibroblast cell line and Hs 700T and MIA PaCa-2 human pancreatic cancer cell lines were stably transfected with MED29 or an empty control vector. ( a ) Relative MED29 mRNA expression levels were assessed by qRT-PCR in MED29 transduced Hs 700T and MIA PaCa-2 cells and their respective empty vector (mock) control cells. MED29 expression values were normalized against a TATA-box binding protein house-keeping gene. ( b ) Western blot was used to detect MED29 protein (21 kDa) in stable MED29-expressing cells vs. mock control cells. β-Tubulin was used as a loading control. ( c ) The growth of the NIH3T3/MED29-1 and NIH3T3/mock cells was monitored at indicated time points in normal growth medium and ( d ) after 24h of serum starvation (1% FBS). ( e ) The growth of the MIAPaCa2/MED29 cells were monitored at indicated time points after release from G1-arrest. The mean +/− s.d. of six replicates are shown. The experiments were repeated three times with similar results. * P

    Article Snippet: Gene expression analyses were performed using the Light Cycler qRT-PCR equipment (Roche Applied Science).

    Techniques: Expressing, In Vitro, Stable Transfection, Transfection, Plasmid Preparation, Quantitative RT-PCR, Binding Assay, Western Blot

    PopRice retrotransposons produce extrachromosomal DNA during seed development in wild type rice. ( A ) Southern blot experiment using non-digested genomic DNA extracted from WT rice leaves and seeds at different stages as indicated and detected with a PopRice specific probe ( gDNA : genomic DNA, ecDNA : extrachromosomal DNA). The GelRed gel picture is shown as a loading control. ( B ) Southern blot experiment using non-digested genomic DNA extracted from dissected rice seed tissues as indicated and detected with a PopRice specific probe. Legend as in A. ( C ) Detection of PopRice circular forms using inverse PCR with primers localization depicted on the right ( black bar : PopRice element, arrows : PCR primers, grey boxes : LTRs). Upper gel: PCR amplification of PopRice circles, middle gel: control PCR for PopRice detection, lower gel: PCR using eEF1α primers as control. ( D ) qRT-PCR analysis of PopRice and Osr4 transcripts in WT rice leaves, flowers and mature seeds. Two pairs of primers were used: PopRice specific primers (top) and primers specific for the whole Osr4 family (including PopRice ) (bottom). The relative expression levels were calculated using eIF-5a as a reference, error bars indicate technical replicates, two biological replicates are shown for each tissue.

    Journal: PLoS Genetics

    Article Title: Sequencing the extrachromosomal circular mobilome reveals retrotransposon activity in plants

    doi: 10.1371/journal.pgen.1006630

    Figure Lengend Snippet: PopRice retrotransposons produce extrachromosomal DNA during seed development in wild type rice. ( A ) Southern blot experiment using non-digested genomic DNA extracted from WT rice leaves and seeds at different stages as indicated and detected with a PopRice specific probe ( gDNA : genomic DNA, ecDNA : extrachromosomal DNA). The GelRed gel picture is shown as a loading control. ( B ) Southern blot experiment using non-digested genomic DNA extracted from dissected rice seed tissues as indicated and detected with a PopRice specific probe. Legend as in A. ( C ) Detection of PopRice circular forms using inverse PCR with primers localization depicted on the right ( black bar : PopRice element, arrows : PCR primers, grey boxes : LTRs). Upper gel: PCR amplification of PopRice circles, middle gel: control PCR for PopRice detection, lower gel: PCR using eEF1α primers as control. ( D ) qRT-PCR analysis of PopRice and Osr4 transcripts in WT rice leaves, flowers and mature seeds. Two pairs of primers were used: PopRice specific primers (top) and primers specific for the whole Osr4 family (including PopRice ) (bottom). The relative expression levels were calculated using eIF-5a as a reference, error bars indicate technical replicates, two biological replicates are shown for each tissue.

    Article Snippet: Analyses by quantitative real-time PCR (qRT-PCR) were established using 7 to 35 ng of cDNA. qRT-PCRs were run on a LightCycler 480 (Roche) using Takyon No Rox SYBR MasterMix dTTP Blue Kit (Eurogentec) according to the manufacturer’s instructions.

    Techniques: Southern Blot, Inverse PCR, Polymerase Chain Reaction, Amplification, Quantitative RT-PCR, Expressing

    Validation of RNA-Seq data by quantitative real-time PCR (qRT-PCR). (A) Comparison of the expressions profile of eight DEGs determined by Illumina HiSeq™ 2000 sequencing platform and qRT-PCR at 48 h (pi) using ribosomal protein subunit S4 ( rps4 ) as housekeeping gene. Data shown are the mean of triplicates ± SD. (B) Correlation of data between RNA-Seq and qRT-PCR techniques.

    Journal: Frontiers in Immunology

    Article Title: Transcriptomic Profiles of Senegalese Sole Infected With Nervous Necrosis Virus Reassortants Presenting Different Degree of Virulence

    doi: 10.3389/fimmu.2018.01626

    Figure Lengend Snippet: Validation of RNA-Seq data by quantitative real-time PCR (qRT-PCR). (A) Comparison of the expressions profile of eight DEGs determined by Illumina HiSeq™ 2000 sequencing platform and qRT-PCR at 48 h (pi) using ribosomal protein subunit S4 ( rps4 ) as housekeeping gene. Data shown are the mean of triplicates ± SD. (B) Correlation of data between RNA-Seq and qRT-PCR techniques.

    Article Snippet: Analysis of Gene Expression by Quantitative Real-Time PCR (qRT-PCR) All the reactions were conducted using the LightCycler 96 Termocycler (Roche) and the Fast Start Essential DNA Green MasterMix (Roche) using SYBR Green technology.

    Techniques: RNA Sequencing Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Sequencing

    Effect of N. arvensis extracts in IB3-1 cells. (A) Effect of N. arvensis extracts on IL-8 mRNA expression in IB3-1 cells. IB3-1 cells were treated with the chloroform extract (solved in EtOH/DMSO 95/5) (A) at different concentrations (0.1–200 μg/ml) for 16 h before infection with PAO1 (100 CFU/cell) for further 4 h. IL-8 mRNA expression was quantified by qRT-PCR. Expression of IL-8 mRNA was measured by Real-Time qPCR and obtained by comparing the ratio IL-8 and the housekeeping gene GAPDH between non-infected and infected cells. The results are expressed as the % of untreated cells. Data are mean ±SEM of three independent experiments performed in duplicate. Dashed line corresponds to cells treated with solvent alone. (B) Effect of N. arvensis extract in PAO1 infected IB3-1 cells after different incubation times. The N. arvensis extract (10 μg/ml) was added to IB3-1 cells 24, 4, and 2 h before, simultaneously or 2 h post PAO1 infection (100 CFU/cell). IL-8 mRNA expression was measured as indicated in (A) . Data are mean ±SEM of three independent experiments performed in duplicate. Dashed line corresponds to cells treated with solvent alone. (C) PAO1 growth. Bacteria were cultured overnight at 37°C in the presence of solvent or ranging doses (0.1–1000 μg/ml) of N. arvensis extract. Bacterial growth was monitored by absorbance measures at 660 nm. A representative experiment performed in duplicate is shown. (D) Adhesion of PAO1 to IB3.1 cells. 500,000 IB3-1 cells on Petri dishes, in duplicate, were treated for 24 h with 10 μg/ml N. arvensis extract. Different amounts of 35 S-PAO1, expressed as CFU/well, were added to the wells and incubated as described in section “Materials and Methods”. Data reported in the figure are the specific binding calculated by subtracting counts obtained in the presence of 100-fold excess of non-labeled PAO1 and are expressed as CFU/well. Data are mean ±SEM of three independent experiments performed in duplicate. (E) Effects of N. arvensis extracts on cell growth in IB3-1 cells. IB3-1 cells were incubated with increasing concentrations (1–200 μg/ml) of the chloroform extract (solved in EtOH/DMSO) of N. arvensis for 24 and 48 h. Cell viability was measured by cytometer analysis. Data are expressed as % control (solvent) and are relative to a representative experiment performed in duplicate. Dashed line corresponds to cells treated with solvent alone.

    Journal: Frontiers in Pharmacology

    Article Title: β-Sitosterol Reduces the Expression of Chemotactic Cytokine Genes in Cystic Fibrosis Bronchial Epithelial Cells

    doi: 10.3389/fphar.2017.00236

    Figure Lengend Snippet: Effect of N. arvensis extracts in IB3-1 cells. (A) Effect of N. arvensis extracts on IL-8 mRNA expression in IB3-1 cells. IB3-1 cells were treated with the chloroform extract (solved in EtOH/DMSO 95/5) (A) at different concentrations (0.1–200 μg/ml) for 16 h before infection with PAO1 (100 CFU/cell) for further 4 h. IL-8 mRNA expression was quantified by qRT-PCR. Expression of IL-8 mRNA was measured by Real-Time qPCR and obtained by comparing the ratio IL-8 and the housekeeping gene GAPDH between non-infected and infected cells. The results are expressed as the % of untreated cells. Data are mean ±SEM of three independent experiments performed in duplicate. Dashed line corresponds to cells treated with solvent alone. (B) Effect of N. arvensis extract in PAO1 infected IB3-1 cells after different incubation times. The N. arvensis extract (10 μg/ml) was added to IB3-1 cells 24, 4, and 2 h before, simultaneously or 2 h post PAO1 infection (100 CFU/cell). IL-8 mRNA expression was measured as indicated in (A) . Data are mean ±SEM of three independent experiments performed in duplicate. Dashed line corresponds to cells treated with solvent alone. (C) PAO1 growth. Bacteria were cultured overnight at 37°C in the presence of solvent or ranging doses (0.1–1000 μg/ml) of N. arvensis extract. Bacterial growth was monitored by absorbance measures at 660 nm. A representative experiment performed in duplicate is shown. (D) Adhesion of PAO1 to IB3.1 cells. 500,000 IB3-1 cells on Petri dishes, in duplicate, were treated for 24 h with 10 μg/ml N. arvensis extract. Different amounts of 35 S-PAO1, expressed as CFU/well, were added to the wells and incubated as described in section “Materials and Methods”. Data reported in the figure are the specific binding calculated by subtracting counts obtained in the presence of 100-fold excess of non-labeled PAO1 and are expressed as CFU/well. Data are mean ±SEM of three independent experiments performed in duplicate. (E) Effects of N. arvensis extracts on cell growth in IB3-1 cells. IB3-1 cells were incubated with increasing concentrations (1–200 μg/ml) of the chloroform extract (solved in EtOH/DMSO) of N. arvensis for 24 and 48 h. Cell viability was measured by cytometer analysis. Data are expressed as % control (solvent) and are relative to a representative experiment performed in duplicate. Dashed line corresponds to cells treated with solvent alone.

    Article Snippet: Quantitative Gene Expression Analyses by qRT-PCR Total RNA from IB3-1 and CuFi-1 cells was purified using a High Pure RNA Isolation Kit (Roche, Mannheim, Germany), and 2.0 μg RNA were reverse transcribed to cDNA using the High Capacity cDNA Archive Kit and random primers (Applied Biosystems, Foster City, CA, USA) in a final reaction volume of 20 μl.

    Techniques: Expressing, Infection, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Incubation, Cell Culture, Binding Assay, Labeling, Cytometry

    Spatial regulation of SA accumulation leading to an organized multicellular response in ETI. (A) Site-specific sampling for SA and PR1 analyses at 7 h.p.i. A nearly complete half of a pPR1-YFP-NLS leaf was fully infiltrated with Pst_a2 (OD 600 = 0.2, upper) or 10 mM MgCl 2 (mock, lower). The leaf was divided into four areas along the mid-rib (numbered 1–4), and three leaf disks (2 mm in diameter) per area were sampled, as shown in a dashed ellipse for zone 1 in the upper right pictures. Representative sample pictures are shown. Scale bars = 2.5 mm. (B) Intensity profiles of YFP and autofluorescence in the white boxes in the Pst_a2 -treated leaf in (A) along the red arrow. (C) Intensity profiles of YFP and autofluorescence in the white boxes in the mock-treated leaf in (A) along the red arrow. (D) The endogenous PR1 expression levels in the four zones were measured by qRT–PCR. Eighteen leaf disks, corresponding to one zone, from six leaves were pooled as one sample. Bars represent means � SD of three biological replicates. (E) The free SA and SA glycoside (SAG) levels in the four zones. Three disks from one zone from one leaf were pooled and analyzed. Bars represent means � SD of three leaves. Experiments were repeated twice with similar results. (F) A schematic summary of an organized concentric pattern of the inner SA and the outer JA active domains which appeared around the infection site of Pst_a2 . (See also Supplementary Fig. S4 ).

    Journal: Plant and Cell Physiology

    Article Title: Salicylic Acid and Jasmonic Acid Pathways are Activated in Spatially Different Domains Around the Infection Site During Effector-Triggered Immunity in Arabidopsis thaliana

    doi: 10.1093/pcp/pcx181

    Figure Lengend Snippet: Spatial regulation of SA accumulation leading to an organized multicellular response in ETI. (A) Site-specific sampling for SA and PR1 analyses at 7 h.p.i. A nearly complete half of a pPR1-YFP-NLS leaf was fully infiltrated with Pst_a2 (OD 600 = 0.2, upper) or 10 mM MgCl 2 (mock, lower). The leaf was divided into four areas along the mid-rib (numbered 1–4), and three leaf disks (2 mm in diameter) per area were sampled, as shown in a dashed ellipse for zone 1 in the upper right pictures. Representative sample pictures are shown. Scale bars = 2.5 mm. (B) Intensity profiles of YFP and autofluorescence in the white boxes in the Pst_a2 -treated leaf in (A) along the red arrow. (C) Intensity profiles of YFP and autofluorescence in the white boxes in the mock-treated leaf in (A) along the red arrow. (D) The endogenous PR1 expression levels in the four zones were measured by qRT–PCR. Eighteen leaf disks, corresponding to one zone, from six leaves were pooled as one sample. Bars represent means � SD of three biological replicates. (E) The free SA and SA glycoside (SAG) levels in the four zones. Three disks from one zone from one leaf were pooled and analyzed. Bars represent means � SD of three leaves. Experiments were repeated twice with similar results. (F) A schematic summary of an organized concentric pattern of the inner SA and the outer JA active domains which appeared around the infection site of Pst_a2 . (See also Supplementary Fig. S4 ).

    Article Snippet: Quantitative reverse transcription–PCR (qRT–PCR) analysis was performed with a LightCycler TaqMan Master (Roche Applied Science) on a LightCycler 480 instrument II (Roche Applied Science).

    Techniques: Sampling, Expressing, Quantitative RT-PCR, Infection

    miR-23a/b promotes the osteogenic differentiation of BMSCs. ( a ) qRT-PCR analysis showed the relative levels of miR-23a/b in BMSCs induced to differentiate into osteoblasts for 14 days. ( b and c ) Representative images of Alizarin Red staining ( b ) and the quantitative analysis of matrix mineralization ( c ) in BMSCs induced to differentiate into osteoblasts for 21 days after transfection. ( d ) ALP activity and osteocalcin secretion were measured in BMSCs induced to generate osteoblasts for 48 h. ( e ) qRT-PCR was used to analyze the relative expression levels of Osterix and Runx2 in BMSCs induced to differentiate into osteoblasts for 48 h. Scale bars: 100 μm. n =5 per group. Data are shown as the mean±s.d. * P

    Journal: Bone Research

    Article Title: miR-23a/b regulates the balance between osteoblast and adipocyte differentiation in bone marrow mesenchymal stem cells

    doi: 10.1038/boneres.2016.22

    Figure Lengend Snippet: miR-23a/b promotes the osteogenic differentiation of BMSCs. ( a ) qRT-PCR analysis showed the relative levels of miR-23a/b in BMSCs induced to differentiate into osteoblasts for 14 days. ( b and c ) Representative images of Alizarin Red staining ( b ) and the quantitative analysis of matrix mineralization ( c ) in BMSCs induced to differentiate into osteoblasts for 21 days after transfection. ( d ) ALP activity and osteocalcin secretion were measured in BMSCs induced to generate osteoblasts for 48 h. ( e ) qRT-PCR was used to analyze the relative expression levels of Osterix and Runx2 in BMSCs induced to differentiate into osteoblasts for 48 h. Scale bars: 100 μm. n =5 per group. Data are shown as the mean±s.d. * P

    Article Snippet: qRT-PCR analysis Quantitative reverse transcription PCR (qRT-PCR) was performed using a Roche Molecular Light Cycler (Basel, Basel-Stadt, Switzerland) as previously described.

    Techniques: Quantitative RT-PCR, Staining, Transfection, ALP Assay, Activity Assay, Expressing

    miR-23a/b directly targets Tmem64 . ( a ) Schematic representation of the predicted miR-23a/b target site in the 3′-UTR of mouse Tmem64 . The alignment of miR-23a/b with WT and MUT 3′-UTR region is shown by complementary pairing, and three mutated nucleotides are underlined. ( b ) BMSCs were co-transfected with the luciferase reporter carrying WT-pGL3- Tmem64 or MUT-pGL3- Tmem64 along with agomiR-23a/b or agomiR-NC. The effects of miR-23a/b on the luciferase reporter constructs were determined 48 h after transfection. The firefly luciferase values were normalized to Renilla luciferase; n =5. ( c ) After BMSCs were transfected with agomiR-23a/b or antagomiR-23a/b, the relative levels of Tmem64 protein expression were determined by western blot; β-actin was used as loading control; n =5. ( d ) The relative levels of Tmem64 mRNA were determined using qRT-PCR and normalized to β-actin; n =5. ( e ) Tmem64 protein levels in BMSCs from 3- and 18-month-old mice were measured by western blot and expressed as the densitometry of Tmem64/β-actin. Tmem64 mRNA levels were determined by qRT-PCR and are shown as the fold-induction relative to β-actin; n =3. ( f ) The increase in ALP activity induced by agomiR-23a/b was blocked by the transfection of MUT Tmem64 3′-UTR into osteogenic-induced-BMSCs. * P

    Journal: Bone Research

    Article Title: miR-23a/b regulates the balance between osteoblast and adipocyte differentiation in bone marrow mesenchymal stem cells

    doi: 10.1038/boneres.2016.22

    Figure Lengend Snippet: miR-23a/b directly targets Tmem64 . ( a ) Schematic representation of the predicted miR-23a/b target site in the 3′-UTR of mouse Tmem64 . The alignment of miR-23a/b with WT and MUT 3′-UTR region is shown by complementary pairing, and three mutated nucleotides are underlined. ( b ) BMSCs were co-transfected with the luciferase reporter carrying WT-pGL3- Tmem64 or MUT-pGL3- Tmem64 along with agomiR-23a/b or agomiR-NC. The effects of miR-23a/b on the luciferase reporter constructs were determined 48 h after transfection. The firefly luciferase values were normalized to Renilla luciferase; n =5. ( c ) After BMSCs were transfected with agomiR-23a/b or antagomiR-23a/b, the relative levels of Tmem64 protein expression were determined by western blot; β-actin was used as loading control; n =5. ( d ) The relative levels of Tmem64 mRNA were determined using qRT-PCR and normalized to β-actin; n =5. ( e ) Tmem64 protein levels in BMSCs from 3- and 18-month-old mice were measured by western blot and expressed as the densitometry of Tmem64/β-actin. Tmem64 mRNA levels were determined by qRT-PCR and are shown as the fold-induction relative to β-actin; n =3. ( f ) The increase in ALP activity induced by agomiR-23a/b was blocked by the transfection of MUT Tmem64 3′-UTR into osteogenic-induced-BMSCs. * P

    Article Snippet: qRT-PCR analysis Quantitative reverse transcription PCR (qRT-PCR) was performed using a Roche Molecular Light Cycler (Basel, Basel-Stadt, Switzerland) as previously described.

    Techniques: Transfection, Luciferase, Construct, Expressing, Western Blot, Quantitative RT-PCR, Mouse Assay, ALP Assay, Activity Assay

    miR-23a/b is gradually downregulated in BMSCs throughout the aging process. ( a ) qRT-PCR was used to analyze the relative levels of miR-23a/b in BMSCs isolated from C57BL/6 mice of different ages. n =5 per group. ( b and c ) Comparison of miR-23a/b levels in young and old human BMSCs as determined by qRT-PCR of human male ( b ) and female samples ( c ). Male: n y =15, n o =17. Female: n y =14, n o =15. Data are shown as the mean±s.d. ** P

    Journal: Bone Research

    Article Title: miR-23a/b regulates the balance between osteoblast and adipocyte differentiation in bone marrow mesenchymal stem cells

    doi: 10.1038/boneres.2016.22

    Figure Lengend Snippet: miR-23a/b is gradually downregulated in BMSCs throughout the aging process. ( a ) qRT-PCR was used to analyze the relative levels of miR-23a/b in BMSCs isolated from C57BL/6 mice of different ages. n =5 per group. ( b and c ) Comparison of miR-23a/b levels in young and old human BMSCs as determined by qRT-PCR of human male ( b ) and female samples ( c ). Male: n y =15, n o =17. Female: n y =14, n o =15. Data are shown as the mean±s.d. ** P

    Article Snippet: qRT-PCR analysis Quantitative reverse transcription PCR (qRT-PCR) was performed using a Roche Molecular Light Cycler (Basel, Basel-Stadt, Switzerland) as previously described.

    Techniques: Quantitative RT-PCR, Isolation, Mouse Assay

    miR-23a/b inhibits the adipogenic differentiation of BMSCs. ( a ) qRT-PCR analysis of the relative levels of miR-23a/b in BMSCs induced to differentiate into adipocytes for 14 days. ( b ) The relative levels of miR-23a/b in BMSCs transfected with 10 μmol·L −1 agomiR-23a/b, antagomiR-23a/b or their NC were analyzed by qRT-PCR. ( c and d ) Representative images of Oil Red staining of lipid droplets ( c ), and the quantitative analysis of the number of Oil Red spots ( d ) in BMSCs induced to differentiate into adipocytes for 14 days. ( e and f ) The relative mRNA expression levels of adipogenic markers, Pparg ( e ) and Fabp4 ( f ), were measured by qRT-PCR in BMSCs induced to differentiate into adipocytes for 48 h. Scale bars: 120 μm. n =5 per group. Data are shown as the mean±s.d. * P

    Journal: Bone Research

    Article Title: miR-23a/b regulates the balance between osteoblast and adipocyte differentiation in bone marrow mesenchymal stem cells

    doi: 10.1038/boneres.2016.22

    Figure Lengend Snippet: miR-23a/b inhibits the adipogenic differentiation of BMSCs. ( a ) qRT-PCR analysis of the relative levels of miR-23a/b in BMSCs induced to differentiate into adipocytes for 14 days. ( b ) The relative levels of miR-23a/b in BMSCs transfected with 10 μmol·L −1 agomiR-23a/b, antagomiR-23a/b or their NC were analyzed by qRT-PCR. ( c and d ) Representative images of Oil Red staining of lipid droplets ( c ), and the quantitative analysis of the number of Oil Red spots ( d ) in BMSCs induced to differentiate into adipocytes for 14 days. ( e and f ) The relative mRNA expression levels of adipogenic markers, Pparg ( e ) and Fabp4 ( f ), were measured by qRT-PCR in BMSCs induced to differentiate into adipocytes for 48 h. Scale bars: 120 μm. n =5 per group. Data are shown as the mean±s.d. * P

    Article Snippet: qRT-PCR analysis Quantitative reverse transcription PCR (qRT-PCR) was performed using a Roche Molecular Light Cycler (Basel, Basel-Stadt, Switzerland) as previously described.

    Techniques: Quantitative RT-PCR, Transfection, Staining, Expressing

    MCF10A cells expressing DCLK1 harbor abnormal chromosomes. A: Whole cell lysates (60 μg of total protein) of parent MCF10A, MCF10A-DCLK1 and MCF10A-DCLK1-K419R cells were immunoblotted by the indicated antibodies. B and C: cDNAs were prepared from parent MCF10A and MCF10A-DCLK1 cells and qRT-PCR was performed for indicated genes. The data were normalized against GAPDH and are shown as mean +/- SD. n.s., not significant; * *, p

    Journal: Biochemistry and Biophysics Reports

    Article Title: Doublecortin-like kinase 1 compromises DNA repair and induces chromosomal instability

    doi: 10.1016/j.bbrep.2018.10.014

    Figure Lengend Snippet: MCF10A cells expressing DCLK1 harbor abnormal chromosomes. A: Whole cell lysates (60 μg of total protein) of parent MCF10A, MCF10A-DCLK1 and MCF10A-DCLK1-K419R cells were immunoblotted by the indicated antibodies. B and C: cDNAs were prepared from parent MCF10A and MCF10A-DCLK1 cells and qRT-PCR was performed for indicated genes. The data were normalized against GAPDH and are shown as mean +/- SD. n.s., not significant; * *, p

    Article Snippet: 2.8 quantitative-RT-PCR (qRT-PCR) qRT-PCR analysis was performed using SYBR Green (Roche) and ABI7500 Real-Time PCR system (Applied Biosystems).

    Techniques: Expressing, Quantitative RT-PCR

    Endogenous ABA content in SpUSP -overexpressing and wild-type 2 month-old plants measured by HLPC. (A) ABA content in well-watered condition. (B) ABA content after withholding water for 7 d. (C) Relative expression level of NCED3 in OE and wild-type (ZS6) lines under drought stress via qRT-PCR. Variance analysis was performed to determine significant differences (* P

    Journal: Journal of Experimental Botany

    Article Title: SpUSP, an annexin-interacting universal stress protein, enhances drought tolerance in tomato

    doi: 10.1093/jxb/ers220

    Figure Lengend Snippet: Endogenous ABA content in SpUSP -overexpressing and wild-type 2 month-old plants measured by HLPC. (A) ABA content in well-watered condition. (B) ABA content after withholding water for 7 d. (C) Relative expression level of NCED3 in OE and wild-type (ZS6) lines under drought stress via qRT-PCR. Variance analysis was performed to determine significant differences (* P

    Article Snippet: Quantitative RT-PCR (qRT-PCR) analysis Quantitative RT-PCR was performed on a LightCycler 480 system (Roche, Switzerland).

    Techniques: Expressing, Quantitative RT-PCR

    Expression patterns of SpUSP. (A) Tissue profiling analysis of SpUSP in different organs of wild tomato LA716 (Solanum pennellii) and cultivated tomato M82 (S. lycopersicum) using qRT-PCR. (B) Expression pattern of SpUSP during a 24h period. Leaf samples were collected every 3h for 24h starting from 06.00h. (C) Expression patterns of SpUSP via GUS staining: (a) leaf, (b) stoma, (c) stem, (d) root, and (e) trichome. (D) Subcellular localization of SpUSP. The photographs were taken under bright light, in the dark field for the GFP-derived green fluorescence and merged respectively. (E) Interaction of SpUSP with annexin via BiFC. The photographs were taken under bright light, in the dark field for YFP-derived green fluorescence, staining with DAPI and overlap, respectively. Scale bars=10 μm.

    Journal: Journal of Experimental Botany

    Article Title: SpUSP, an annexin-interacting universal stress protein, enhances drought tolerance in tomato

    doi: 10.1093/jxb/ers220

    Figure Lengend Snippet: Expression patterns of SpUSP. (A) Tissue profiling analysis of SpUSP in different organs of wild tomato LA716 (Solanum pennellii) and cultivated tomato M82 (S. lycopersicum) using qRT-PCR. (B) Expression pattern of SpUSP during a 24h period. Leaf samples were collected every 3h for 24h starting from 06.00h. (C) Expression patterns of SpUSP via GUS staining: (a) leaf, (b) stoma, (c) stem, (d) root, and (e) trichome. (D) Subcellular localization of SpUSP. The photographs were taken under bright light, in the dark field for the GFP-derived green fluorescence and merged respectively. (E) Interaction of SpUSP with annexin via BiFC. The photographs were taken under bright light, in the dark field for YFP-derived green fluorescence, staining with DAPI and overlap, respectively. Scale bars=10 μm.

    Article Snippet: Quantitative RT-PCR (qRT-PCR) analysis Quantitative RT-PCR was performed on a LightCycler 480 system (Roche, Switzerland).

    Techniques: Expressing, Quantitative RT-PCR, Staining, Derivative Assay, Fluorescence, Bimolecular Fluorescence Complementation Assay

    SpUSP overexpression enhances drought tolerance in tomato. (A) Drought tolerance tests for SpSUP -overexpressing (OE44, OE54, and OE69) and wild-type ZS6 plants grown in the same pot. The phenotypes under well-watered (‘Control’) and drought-stress conditions (‘Drought’) are shown.. (B) Effects of drought on the fresh and dry weights of the OE and wild-type lines. (C) Chlorophyll contents in plant leaves under drought stress (‘DS’) or normal condition (‘CK’). (D) Proline accumulation in plant leaves under drought stress. (E) Relative expression levels of P5CS1 in the OE and wild-type lines under drought stress via qRT-PCR. (F) Soluble sugar content in plant leaves under drought stress. (G) MDA content in plant leaves under stress (‘Drought’) or normal condition (‘Control’). The data shown are the mean ±SE ( n =3). Single (* P

    Journal: Journal of Experimental Botany

    Article Title: SpUSP, an annexin-interacting universal stress protein, enhances drought tolerance in tomato

    doi: 10.1093/jxb/ers220

    Figure Lengend Snippet: SpUSP overexpression enhances drought tolerance in tomato. (A) Drought tolerance tests for SpSUP -overexpressing (OE44, OE54, and OE69) and wild-type ZS6 plants grown in the same pot. The phenotypes under well-watered (‘Control’) and drought-stress conditions (‘Drought’) are shown.. (B) Effects of drought on the fresh and dry weights of the OE and wild-type lines. (C) Chlorophyll contents in plant leaves under drought stress (‘DS’) or normal condition (‘CK’). (D) Proline accumulation in plant leaves under drought stress. (E) Relative expression levels of P5CS1 in the OE and wild-type lines under drought stress via qRT-PCR. (F) Soluble sugar content in plant leaves under drought stress. (G) MDA content in plant leaves under stress (‘Drought’) or normal condition (‘Control’). The data shown are the mean ±SE ( n =3). Single (* P

    Article Snippet: Quantitative RT-PCR (qRT-PCR) analysis Quantitative RT-PCR was performed on a LightCycler 480 system (Roche, Switzerland).

    Techniques: Over Expression, Expressing, Quantitative RT-PCR, Multiple Displacement Amplification

    Growth performances of OE SpUSP and wild-type seedlings treated with mannitol, salt, and ABA stresses. (A) Analysis of SpUSP transcriptional expression via qRT-PCR in overexpressing (OE44, OE54, and OE69) and wild-type (ZS6) lines. Seedling lengths and weights of transgenic and wild-type lines after treatment with 200mM mannitol (B), 100mM NaCl (C), and 3 μM ABA (D), and without stress as a control. The seedlings were grown in half-strength MS medium. The data shown are the mean ±SE ( n = 6). Single (* P

    Journal: Journal of Experimental Botany

    Article Title: SpUSP, an annexin-interacting universal stress protein, enhances drought tolerance in tomato

    doi: 10.1093/jxb/ers220

    Figure Lengend Snippet: Growth performances of OE SpUSP and wild-type seedlings treated with mannitol, salt, and ABA stresses. (A) Analysis of SpUSP transcriptional expression via qRT-PCR in overexpressing (OE44, OE54, and OE69) and wild-type (ZS6) lines. Seedling lengths and weights of transgenic and wild-type lines after treatment with 200mM mannitol (B), 100mM NaCl (C), and 3 μM ABA (D), and without stress as a control. The seedlings were grown in half-strength MS medium. The data shown are the mean ±SE ( n = 6). Single (* P

    Article Snippet: Quantitative RT-PCR (qRT-PCR) analysis Quantitative RT-PCR was performed on a LightCycler 480 system (Roche, Switzerland).

    Techniques: Expressing, Quantitative RT-PCR, Transgenic Assay, Mass Spectrometry

    si-cJun and sh-cJun activity in melanoma after AGO2 re-expression. ( A ) cJun-qRT–PCR analysis of Mel-Ju cells after 2 μ g pAGO2 or 2 μ g pIRES (mock) transfection and si-cJun treatment (cJun inhibition per pmol si-cJun). cJun inhibition increases after AGO2 re-expression up to 70% compared with mock. ( B ) cJun qRT–PCR (* P

    Journal: British Journal of Cancer

    Article Title: Strong reduction of AGO2 expression in melanoma and cellular consequences

    doi: 10.1038/bjc.2013.646

    Figure Lengend Snippet: si-cJun and sh-cJun activity in melanoma after AGO2 re-expression. ( A ) cJun-qRT–PCR analysis of Mel-Ju cells after 2 μ g pAGO2 or 2 μ g pIRES (mock) transfection and si-cJun treatment (cJun inhibition per pmol si-cJun). cJun inhibition increases after AGO2 re-expression up to 70% compared with mock. ( B ) cJun qRT–PCR (* P

    Article Snippet: Analysis of gene expression by quantitative PCR Quantitative real time-PCR (qRT–PCR) was performed on a Lightcycler (Roche, Mannheim, Germany). cDNA template (500 ng), 0.5 μ l (20 μ M ) of forward and reverse primers and 10 μ l of Sybr Premix Ex Taq (TaKaRa, Shiga, Japan) in a total volume of 20 μ l were applied to the following PCR program: 30 s 95 °C (initial denaturation); 20 °C s−1 temperature transition rate up to 95 °C for 15 s, 10 s annealing, 20 s 72 °C, 10 s acquisition mode single, repeated for 45 times (amplification).

    Techniques: Activity Assay, Expressing, Quantitative RT-PCR, Transfection, Inhibition