quantitative polymerase chain reaction qpcr  (TaKaRa)

 
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    Structured Review

    TaKaRa quantitative polymerase chain reaction qpcr
    Rapamycin promotes SBLC transcription through effects on E2F1. ( A ) HCT116 and A549 cells were treated with rapamycin, metformin, resveratrol and spermidine (1 μM, 1 mM, 25 μM and 0.1 μM, respectively) for 24 h before measuring the SBLC levels by <t>qPCR.</t> ( B ) SBLC levels in HCT116 cells measured by qPCR (left) along with the levels of c-Myc and p21 by blotting (right) after addition of 1 μM rapamycin. β-actin was included throughout as a loading control. ( C ) The experiment in (B) was repeated using A549 cells. ( D ) Schematic of predicted TF binding sites for E2F1, USF-1, MAZ, SP1 and CREB in SBLC . ( E ) A549 and HCT116 cells were treated with 1 μM rapamycin and the levels of TFs in (D) were measured by western blot. Relative levels of E2F1 were estimated using densitometric analysis. ( F ) Rapamycin was added to HCT116 cells for 24 h in which E2F1 had been depleted by shRNA along with respective controls. SBLC levels measured by qPCR (top) along with E2F1 by blotting (bottom). ( G ) The experiment in (F) was repeated on HCT116 cells after treatment with sh-SBLC. ( H ) Localization of the three intergenic E2F1 binding sites in SBLC (left). ChIP using E2F1 or IgG control assessed by <t>RT-PCR</t> analysis against B1 and B2 regions (right). (A–H) Results are representative of three independent experiments. Values are mean ± SEM. NS, not significant, * P
    Quantitative Polymerase Chain Reaction Qpcr, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "SENEBLOC, a long non-coding RNA suppresses senescence via p53-dependent and independent mechanisms"

    Article Title: SENEBLOC, a long non-coding RNA suppresses senescence via p53-dependent and independent mechanisms

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkaa063

    Rapamycin promotes SBLC transcription through effects on E2F1. ( A ) HCT116 and A549 cells were treated with rapamycin, metformin, resveratrol and spermidine (1 μM, 1 mM, 25 μM and 0.1 μM, respectively) for 24 h before measuring the SBLC levels by qPCR. ( B ) SBLC levels in HCT116 cells measured by qPCR (left) along with the levels of c-Myc and p21 by blotting (right) after addition of 1 μM rapamycin. β-actin was included throughout as a loading control. ( C ) The experiment in (B) was repeated using A549 cells. ( D ) Schematic of predicted TF binding sites for E2F1, USF-1, MAZ, SP1 and CREB in SBLC . ( E ) A549 and HCT116 cells were treated with 1 μM rapamycin and the levels of TFs in (D) were measured by western blot. Relative levels of E2F1 were estimated using densitometric analysis. ( F ) Rapamycin was added to HCT116 cells for 24 h in which E2F1 had been depleted by shRNA along with respective controls. SBLC levels measured by qPCR (top) along with E2F1 by blotting (bottom). ( G ) The experiment in (F) was repeated on HCT116 cells after treatment with sh-SBLC. ( H ) Localization of the three intergenic E2F1 binding sites in SBLC (left). ChIP using E2F1 or IgG control assessed by RT-PCR analysis against B1 and B2 regions (right). (A–H) Results are representative of three independent experiments. Values are mean ± SEM. NS, not significant, * P
    Figure Legend Snippet: Rapamycin promotes SBLC transcription through effects on E2F1. ( A ) HCT116 and A549 cells were treated with rapamycin, metformin, resveratrol and spermidine (1 μM, 1 mM, 25 μM and 0.1 μM, respectively) for 24 h before measuring the SBLC levels by qPCR. ( B ) SBLC levels in HCT116 cells measured by qPCR (left) along with the levels of c-Myc and p21 by blotting (right) after addition of 1 μM rapamycin. β-actin was included throughout as a loading control. ( C ) The experiment in (B) was repeated using A549 cells. ( D ) Schematic of predicted TF binding sites for E2F1, USF-1, MAZ, SP1 and CREB in SBLC . ( E ) A549 and HCT116 cells were treated with 1 μM rapamycin and the levels of TFs in (D) were measured by western blot. Relative levels of E2F1 were estimated using densitometric analysis. ( F ) Rapamycin was added to HCT116 cells for 24 h in which E2F1 had been depleted by shRNA along with respective controls. SBLC levels measured by qPCR (top) along with E2F1 by blotting (bottom). ( G ) The experiment in (F) was repeated on HCT116 cells after treatment with sh-SBLC. ( H ) Localization of the three intergenic E2F1 binding sites in SBLC (left). ChIP using E2F1 or IgG control assessed by RT-PCR analysis against B1 and B2 regions (right). (A–H) Results are representative of three independent experiments. Values are mean ± SEM. NS, not significant, * P

    Techniques Used: Real-time Polymerase Chain Reaction, Binding Assay, Western Blot, shRNA, Chromatin Immunoprecipitation, Reverse Transcription Polymerase Chain Reaction

    Related Articles

    Real-time Polymerase Chain Reaction:

    Article Title: Context-dependent regulation of endothelial cell metabolism: differential effects of the PPARβ/δ agonist GW0742 and VEGF-A
    Article Snippet: .. Quantitative PCR (qPCR), using both intercalator (SYBR® Premix Ex Taq™; Clontech: TaKara Bio) and probe-based (TaqManTM ) technologies, was performed to determine changes in gene expression. .. CPT1A (Hs00912671_m1) and PPARβ/δ (Hs00606407_m1) TaqManTM primer-probes were obtained from Applied Biosystems (ThermoFisher Scientific; UK).

    Article Title: SENEBLOC, a long non-coding RNA suppresses senescence via p53-dependent and independent mechanisms
    Article Snippet: .. Quantitative polymerase chain reaction (qPCR) was performed using SYBR Green real‐time PCR analysis (Takara) with the specified primers ( ). .. PCR results, recorded as cycle threshold (Ct), were normalized against an internal control (β‐actin).

    Article Title: Reactive fibrosis precedes doxorubicin‐induced heart failure through sterile inflammation) Reactive fibrosis precedes doxorubicin‐induced heart failure through sterile inflammation
    Article Snippet: .. Quantitative polymerase chain reaction (qPCR) was prepared using SYBR Fast qPCR Mix (TaKaRa Bio). .. Reverse transcription–polymerase chain reaction was performed on the StepOnePlus Real‐Time PCR System (Applied Biosystems, CA, USA).

    Article Title: Integrated Hypothalamic Transcriptome Profiling Reveals the Reproductive Roles of mRNAs and miRNAs in Sheep
    Article Snippet: .. Furthermore, quantitative PCR (qPCR) was conducted with the SYBR Green qPCR Mix (TaKaRa, Dalian, China) for mRNAs and miRcute Plus miRNA qPCR Kit (TIANGEN, Beijing, China) for miRNAs using a RocheLight Cycler® 480 II system (Roche Applied Science, Mannheim, Germany). .. In addition, β-actin (for mRNA) and U6 small nuclear RNA (snRNA; for miRNA) were utilized as reference gene/miRNA to calculate the relative expression level with the method of 2-ΔΔct ( ).

    Article Title: Akt inhibition synergizes with polycomb repressive complex 2 inhibition in the treatment of multiple myeloma, et al. Akt inhibition synergizes with polycomb repressive complex 2 inhibition in the treatment of multiple myeloma
    Article Snippet: .. In the ChIP assay, quantitative PCR (qPCR) was performed with the Step One Plus real‐time PCR system using SYBR Premix ExTaq II (Tli RNase Plus) from Takara. ..

    Article Title: AID and TET2 co‐operation modulates FANCA expression by active demethylation in diffuse large B cell lymphoma
    Article Snippet: .. After reversal of cross‐links and purification of the DNA, quantitative PCR (qPCR) was performed using SYBR Premix Ex TaqTM (TaKaRa) with Mx3000 thermocycler (Agilent Technologies). .. The primer sequences used for qPCR are listed in the Supporting information, Table .

    Article Title: Pinocembrin alleviates ulcerative colitis in mice via regulating gut microbiota, suppressing TLR4/MD2/NF-κB pathway and promoting intestinal barrier
    Article Snippet: .. All the Quantitative polymerase chain reaction (qPCR) reagents and Reverse Transcriptase kit were from Takara Biotechnology (Shiga, Japan). .. Cell counting kit 8 (CCK-8) assay kit was from Dalian Meilun Biological Technology Co., Ltd. (Dalian, China).

    Article Title: Activation of PXR by alantolactone ameliorates DSS-induced experimental colitis via suppressing NF-κB signaling pathway
    Article Snippet: .. Quantitative polymerase chain reaction (qPCR) was carried out using SYBR Premix ExTaq Mix (Takara Bio Inc., Otsu, Japan) and quantitatively measured with an ABI Prism 7900HT Sequence Detection System (Life Technologies,s, Carlsbad, CA). .. The relative expression of mRNA was normalized as the ratio of optimal density relative to β-actin.

    Purification:

    Article Title: AID and TET2 co‐operation modulates FANCA expression by active demethylation in diffuse large B cell lymphoma
    Article Snippet: .. After reversal of cross‐links and purification of the DNA, quantitative PCR (qPCR) was performed using SYBR Premix Ex TaqTM (TaKaRa) with Mx3000 thermocycler (Agilent Technologies). .. The primer sequences used for qPCR are listed in the Supporting information, Table .

    SYBR Green Assay:

    Article Title: SENEBLOC, a long non-coding RNA suppresses senescence via p53-dependent and independent mechanisms
    Article Snippet: .. Quantitative polymerase chain reaction (qPCR) was performed using SYBR Green real‐time PCR analysis (Takara) with the specified primers ( ). .. PCR results, recorded as cycle threshold (Ct), were normalized against an internal control (β‐actin).

    Article Title: Integrated Hypothalamic Transcriptome Profiling Reveals the Reproductive Roles of mRNAs and miRNAs in Sheep
    Article Snippet: .. Furthermore, quantitative PCR (qPCR) was conducted with the SYBR Green qPCR Mix (TaKaRa, Dalian, China) for mRNAs and miRcute Plus miRNA qPCR Kit (TIANGEN, Beijing, China) for miRNAs using a RocheLight Cycler® 480 II system (Roche Applied Science, Mannheim, Germany). .. In addition, β-actin (for mRNA) and U6 small nuclear RNA (snRNA; for miRNA) were utilized as reference gene/miRNA to calculate the relative expression level with the method of 2-ΔΔct ( ).

    Expressing:

    Article Title: Context-dependent regulation of endothelial cell metabolism: differential effects of the PPARβ/δ agonist GW0742 and VEGF-A
    Article Snippet: .. Quantitative PCR (qPCR), using both intercalator (SYBR® Premix Ex Taq™; Clontech: TaKara Bio) and probe-based (TaqManTM ) technologies, was performed to determine changes in gene expression. .. CPT1A (Hs00912671_m1) and PPARβ/δ (Hs00606407_m1) TaqManTM primer-probes were obtained from Applied Biosystems (ThermoFisher Scientific; UK).

    Sequencing:

    Article Title: Activation of PXR by alantolactone ameliorates DSS-induced experimental colitis via suppressing NF-κB signaling pathway
    Article Snippet: .. Quantitative polymerase chain reaction (qPCR) was carried out using SYBR Premix ExTaq Mix (Takara Bio Inc., Otsu, Japan) and quantitatively measured with an ABI Prism 7900HT Sequence Detection System (Life Technologies,s, Carlsbad, CA). .. The relative expression of mRNA was normalized as the ratio of optimal density relative to β-actin.

    Chromatin Immunoprecipitation:

    Article Title: Akt inhibition synergizes with polycomb repressive complex 2 inhibition in the treatment of multiple myeloma, et al. Akt inhibition synergizes with polycomb repressive complex 2 inhibition in the treatment of multiple myeloma
    Article Snippet: .. In the ChIP assay, quantitative PCR (qPCR) was performed with the Step One Plus real‐time PCR system using SYBR Premix ExTaq II (Tli RNase Plus) from Takara. ..

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    TaKaRa real time quantitative polymerase chain reaction qrt pcr total rna
    LncRNA CASC2 inhibited hypoxia-induced cell proliferation in rat pulmonary arterial tissues. ( a ) The mRNA expression of PCNA was detected by <t>qRT-PCR</t> in rats′ PAs tissues. ( b , c ) The protein expression of PCNA was detected by western blot analysis in rat pulmonary arterial tissues. ## p
    Real Time Quantitative Polymerase Chain Reaction Qrt Pcr Total Rna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa real time quantitative pcr
    Real-time quantitative <t>PCR</t> analysis of IL-4, IL-13, IL-6, IL-12p35, IL-12p40, and IFNγ mRNA levels in splenocytes isolated from OVA-immunized mice that were subsequently treated with 10 μg/mL OVA + 3.0 μM ODN 1612 , ODN 1555 , A151, H154, or iSG3. Mouse splenocytes were pre-incubated in medium for 3 h prior to exposure to OVA and ODNs for 72 h. Expression of IL-4 (A), IL-13 (B), IL-6 (C), IL-12p35 (D), IL-12p40 (E), and IFNγ (F) mRNAs were determined with real-time quantitative PCR. Results are shown as the ratio of <t>cytokine</t> mRNA levels (normalized to β-actin; see Section 2 ) in stimulated versus non-treated cells, as well as the level in the medium control. Values represent means, and error bars indicate the SD of three independent experiments. Values with different letters ( i.e., a, b, c, d, and e) were significantly different ( P
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    99
    TaKaRa premix ex taq reagent
    Hh-dependent induction of osteogenic marker ALP in patient-derived iPSCs. (A) Control iPSCs (KD iPSCs, 201B7iPS cells, and Nips B2 iPSCs) and patient-derived iPSCs (G-OF1iPS1, and G-OF2iPS5) were cultured in OBM, OBM with FIT (bFGF, IGF-1, and TGF-β1), or OBM with SAG. FIT- and SAG-induced ALP activity significantly increased in patient-derived iPSCs compared with control iPSCs. (B) A sample of each supernatant was transferred to tubes for measurement of ALP activity as the hydrolysis of ρ-nitrophenyl phosphate using a LabAssay ALP kit and protein concentration using a Protein assay rapid kit. G-OF1iPS1 was showed a high ALP activity compared with 201B7,especially cultured in OBM with SAG. (C) KDiPSCs and G-OF1iPS1 cells were cultured in OBM (control), or OBM supplemented with 100 ng/ml BMP2/7 for 24 h. qRT-PCR analysis of <t>RUNX2</t> was conducted using Premix Ex <t>Taq</t> reagent according to the manufacturer’s instructions. RUNX2 levels were normalized to those of GAPDH. A significant difference was observed in G-OF1iPS1. Especially, BMP2/7-induced RUNX2 expression promoted an approximate 2-fold increase in the number of G-OFiPSCs compared with KDiPSCs. Values represent the mean ± SD. Bonferroni correction for multiple comparisons was applied, * p
    Premix Ex Taq Reagent, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa one step rt pcr kit
    PGRN deficiency increased the production of systemic and local pro‐inflammatory mediators after injection of LPS . ELISA of serum levels of inflammatory mediators including tumour necrosis factor α ( TNF ‐α) ( A ), interleukin 6 ( IL ‐6) ( B ), IL ‐10 ( C ) and monocyte chemoattractant protein 1 ( MCP ‐1) ( D ). ( E ) Western blot analysis of cold‐inducible <t>RNA</t> ‐binding protein ( CIRP ) and high mobility group box protein 1 ( HMGB 1) in the serum from WT and Grn −/− mice with LPS injection at 6 and 24 hrs. PS red, Ponceau S red staining. Real‐time RT ‐ <t>PCR</t> analysis of mRNA levels of TNF ‐α ( F ), IL ‐6 ( G ), IL ‐1β ( H ) and IL ‐10 ( I ) in the lung after LPS injection. Data are mean ± S.D. * P
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    LncRNA CASC2 inhibited hypoxia-induced cell proliferation in rat pulmonary arterial tissues. ( a ) The mRNA expression of PCNA was detected by qRT-PCR in rats′ PAs tissues. ( b , c ) The protein expression of PCNA was detected by western blot analysis in rat pulmonary arterial tissues. ## p

    Journal: Respiratory Research

    Article Title: Long non-coding RNA CASC2 suppresses pulmonary artery smooth muscle cell proliferation and phenotypic switch in hypoxia-induced pulmonary hypertension

    doi: 10.1186/s12931-019-1018-x

    Figure Lengend Snippet: LncRNA CASC2 inhibited hypoxia-induced cell proliferation in rat pulmonary arterial tissues. ( a ) The mRNA expression of PCNA was detected by qRT-PCR in rats′ PAs tissues. ( b , c ) The protein expression of PCNA was detected by western blot analysis in rat pulmonary arterial tissues. ## p

    Article Snippet: Real time-quantitative polymerase chain reaction (qRT-PCR) Total RNA from PASMCs or rat pulmonary arterial tissues was extracted using the Trizol reagent and reversely transcribed into cDNA using Prime-Script RT-PCR kit (Takara, Dalian, China) following the manufacturer′s instructions.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot

    Up-regulation of lncRNA CASC2 inhibited proliferation in hypoxia-induced PASMCs. ( a ) The expression of lncRNA CASC2 was detected by qRT-PCR in PASMCs. ( b ) Cell number of PASMCs was counted by CyQuant cell proliferation assay kit. ( c ) The mRNA expression of PCNA was detected by qRT-PCR in PASMCs. ( d ) The protein expression of PCNA was detected by western blot analysis in PASMCs. ( e , f ) Cell proliferation status of PASMCs was assessed by EdU staining assay. ### p

    Journal: Respiratory Research

    Article Title: Long non-coding RNA CASC2 suppresses pulmonary artery smooth muscle cell proliferation and phenotypic switch in hypoxia-induced pulmonary hypertension

    doi: 10.1186/s12931-019-1018-x

    Figure Lengend Snippet: Up-regulation of lncRNA CASC2 inhibited proliferation in hypoxia-induced PASMCs. ( a ) The expression of lncRNA CASC2 was detected by qRT-PCR in PASMCs. ( b ) Cell number of PASMCs was counted by CyQuant cell proliferation assay kit. ( c ) The mRNA expression of PCNA was detected by qRT-PCR in PASMCs. ( d ) The protein expression of PCNA was detected by western blot analysis in PASMCs. ( e , f ) Cell proliferation status of PASMCs was assessed by EdU staining assay. ### p

    Article Snippet: Real time-quantitative polymerase chain reaction (qRT-PCR) Total RNA from PASMCs or rat pulmonary arterial tissues was extracted using the Trizol reagent and reversely transcribed into cDNA using Prime-Script RT-PCR kit (Takara, Dalian, China) following the manufacturer′s instructions.

    Techniques: Expressing, Quantitative RT-PCR, CyQUANT Assay, Proliferation Assay, Western Blot, Staining

    LncRNA CASC2 was down-regulated in hypoxia-induced rats and reversed the increase of mPAP and progression of right ventricular hypertrophy caused by hypoxia. ( a ) The expression of lncRNA CASC2 was detected by qRT-PCR in rat pulmonary arterial tissues. ( b ) Systemic blood pressure (SBP) was assessed in normoxia and hypoxia-induced rats. ( c ) Mean pulmonary arterial pressure (mPAP) was detected in normoxia and hypoxia-induced rats. ( d ) The right ventricular hypertrophy index (RVI) was calculated in normoxia and hypoxia-induced rats. # p

    Journal: Respiratory Research

    Article Title: Long non-coding RNA CASC2 suppresses pulmonary artery smooth muscle cell proliferation and phenotypic switch in hypoxia-induced pulmonary hypertension

    doi: 10.1186/s12931-019-1018-x

    Figure Lengend Snippet: LncRNA CASC2 was down-regulated in hypoxia-induced rats and reversed the increase of mPAP and progression of right ventricular hypertrophy caused by hypoxia. ( a ) The expression of lncRNA CASC2 was detected by qRT-PCR in rat pulmonary arterial tissues. ( b ) Systemic blood pressure (SBP) was assessed in normoxia and hypoxia-induced rats. ( c ) Mean pulmonary arterial pressure (mPAP) was detected in normoxia and hypoxia-induced rats. ( d ) The right ventricular hypertrophy index (RVI) was calculated in normoxia and hypoxia-induced rats. # p

    Article Snippet: Real time-quantitative polymerase chain reaction (qRT-PCR) Total RNA from PASMCs or rat pulmonary arterial tissues was extracted using the Trizol reagent and reversely transcribed into cDNA using Prime-Script RT-PCR kit (Takara, Dalian, China) following the manufacturer′s instructions.

    Techniques: Expressing, Quantitative RT-PCR

    Real-time quantitative PCR analysis of IL-4, IL-13, IL-6, IL-12p35, IL-12p40, and IFNγ mRNA levels in splenocytes isolated from OVA-immunized mice that were subsequently treated with 10 μg/mL OVA + 3.0 μM ODN 1612 , ODN 1555 , A151, H154, or iSG3. Mouse splenocytes were pre-incubated in medium for 3 h prior to exposure to OVA and ODNs for 72 h. Expression of IL-4 (A), IL-13 (B), IL-6 (C), IL-12p35 (D), IL-12p40 (E), and IFNγ (F) mRNAs were determined with real-time quantitative PCR. Results are shown as the ratio of cytokine mRNA levels (normalized to β-actin; see Section 2 ) in stimulated versus non-treated cells, as well as the level in the medium control. Values represent means, and error bars indicate the SD of three independent experiments. Values with different letters ( i.e., a, b, c, d, and e) were significantly different ( P

    Journal: FEBS Open Bio

    Article Title: Class I/II hybrid inhibitory oligodeoxynucleotide exerts Th1 and Th2 double immunosuppression

    doi: 10.1016/j.fob.2012.11.002

    Figure Lengend Snippet: Real-time quantitative PCR analysis of IL-4, IL-13, IL-6, IL-12p35, IL-12p40, and IFNγ mRNA levels in splenocytes isolated from OVA-immunized mice that were subsequently treated with 10 μg/mL OVA + 3.0 μM ODN 1612 , ODN 1555 , A151, H154, or iSG3. Mouse splenocytes were pre-incubated in medium for 3 h prior to exposure to OVA and ODNs for 72 h. Expression of IL-4 (A), IL-13 (B), IL-6 (C), IL-12p35 (D), IL-12p40 (E), and IFNγ (F) mRNAs were determined with real-time quantitative PCR. Results are shown as the ratio of cytokine mRNA levels (normalized to β-actin; see Section 2 ) in stimulated versus non-treated cells, as well as the level in the medium control. Values represent means, and error bars indicate the SD of three independent experiments. Values with different letters ( i.e., a, b, c, d, and e) were significantly different ( P

    Article Snippet: An equivalent volume of cDNA was used for quantification of various cytokine cDNAs with real-time quantitative PCR using a Thermal Cycler Dice® Real Time System (TaKaRa Bio Inc., Tokyo, Japan).

    Techniques: Real-time Polymerase Chain Reaction, Isolation, Mouse Assay, Incubation, Expressing

    Hh-dependent induction of osteogenic marker ALP in patient-derived iPSCs. (A) Control iPSCs (KD iPSCs, 201B7iPS cells, and Nips B2 iPSCs) and patient-derived iPSCs (G-OF1iPS1, and G-OF2iPS5) were cultured in OBM, OBM with FIT (bFGF, IGF-1, and TGF-β1), or OBM with SAG. FIT- and SAG-induced ALP activity significantly increased in patient-derived iPSCs compared with control iPSCs. (B) A sample of each supernatant was transferred to tubes for measurement of ALP activity as the hydrolysis of ρ-nitrophenyl phosphate using a LabAssay ALP kit and protein concentration using a Protein assay rapid kit. G-OF1iPS1 was showed a high ALP activity compared with 201B7,especially cultured in OBM with SAG. (C) KDiPSCs and G-OF1iPS1 cells were cultured in OBM (control), or OBM supplemented with 100 ng/ml BMP2/7 for 24 h. qRT-PCR analysis of RUNX2 was conducted using Premix Ex Taq reagent according to the manufacturer’s instructions. RUNX2 levels were normalized to those of GAPDH. A significant difference was observed in G-OF1iPS1. Especially, BMP2/7-induced RUNX2 expression promoted an approximate 2-fold increase in the number of G-OFiPSCs compared with KDiPSCs. Values represent the mean ± SD. Bonferroni correction for multiple comparisons was applied, * p

    Journal: PLoS ONE

    Article Title: Gorlin syndrome-derived induced pluripotent stem cells are hypersensitive to hedgehog-mediated osteogenic induction

    doi: 10.1371/journal.pone.0186879

    Figure Lengend Snippet: Hh-dependent induction of osteogenic marker ALP in patient-derived iPSCs. (A) Control iPSCs (KD iPSCs, 201B7iPS cells, and Nips B2 iPSCs) and patient-derived iPSCs (G-OF1iPS1, and G-OF2iPS5) were cultured in OBM, OBM with FIT (bFGF, IGF-1, and TGF-β1), or OBM with SAG. FIT- and SAG-induced ALP activity significantly increased in patient-derived iPSCs compared with control iPSCs. (B) A sample of each supernatant was transferred to tubes for measurement of ALP activity as the hydrolysis of ρ-nitrophenyl phosphate using a LabAssay ALP kit and protein concentration using a Protein assay rapid kit. G-OF1iPS1 was showed a high ALP activity compared with 201B7,especially cultured in OBM with SAG. (C) KDiPSCs and G-OF1iPS1 cells were cultured in OBM (control), or OBM supplemented with 100 ng/ml BMP2/7 for 24 h. qRT-PCR analysis of RUNX2 was conducted using Premix Ex Taq reagent according to the manufacturer’s instructions. RUNX2 levels were normalized to those of GAPDH. A significant difference was observed in G-OF1iPS1. Especially, BMP2/7-induced RUNX2 expression promoted an approximate 2-fold increase in the number of G-OFiPSCs compared with KDiPSCs. Values represent the mean ± SD. Bonferroni correction for multiple comparisons was applied, * p

    Article Snippet: Quantitative real-time RT-PCR (qRT-PCR) analysis of GLI family zinc finger 1 (GLI1 ) and Runt-related transcription factor 2 (RUNX2) was conducted using Premix Ex Taq reagent (Takara Bio Inc., Shiga, Japan), according to the manufacturer’s instructions.

    Techniques: Marker, ALP Assay, Derivative Assay, Cell Culture, Activity Assay, Protein Concentration, Quantitative RT-PCR, Expressing

    PGRN deficiency increased the production of systemic and local pro‐inflammatory mediators after injection of LPS . ELISA of serum levels of inflammatory mediators including tumour necrosis factor α ( TNF ‐α) ( A ), interleukin 6 ( IL ‐6) ( B ), IL ‐10 ( C ) and monocyte chemoattractant protein 1 ( MCP ‐1) ( D ). ( E ) Western blot analysis of cold‐inducible RNA ‐binding protein ( CIRP ) and high mobility group box protein 1 ( HMGB 1) in the serum from WT and Grn −/− mice with LPS injection at 6 and 24 hrs. PS red, Ponceau S red staining. Real‐time RT ‐ PCR analysis of mRNA levels of TNF ‐α ( F ), IL ‐6 ( G ), IL ‐1β ( H ) and IL ‐10 ( I ) in the lung after LPS injection. Data are mean ± S.D. * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Progranulin deficiency leads to severe inflammation, lung injury and cell death in a mouse model of endotoxic shock

    doi: 10.1111/jcmm.12756

    Figure Lengend Snippet: PGRN deficiency increased the production of systemic and local pro‐inflammatory mediators after injection of LPS . ELISA of serum levels of inflammatory mediators including tumour necrosis factor α ( TNF ‐α) ( A ), interleukin 6 ( IL ‐6) ( B ), IL ‐10 ( C ) and monocyte chemoattractant protein 1 ( MCP ‐1) ( D ). ( E ) Western blot analysis of cold‐inducible RNA ‐binding protein ( CIRP ) and high mobility group box protein 1 ( HMGB 1) in the serum from WT and Grn −/− mice with LPS injection at 6 and 24 hrs. PS red, Ponceau S red staining. Real‐time RT ‐ PCR analysis of mRNA levels of TNF ‐α ( F ), IL ‐6 ( G ), IL ‐1β ( H ) and IL ‐10 ( I ) in the lung after LPS injection. Data are mean ± S.D. * P

    Article Snippet: Then, 1 μg of DNA‐free total RNA was reverse transcribed by use of a one‐step RT‐PCR kit (TaKaRa Bio, Shiga, Japan).

    Techniques: Injection, Enzyme-linked Immunosorbent Assay, Western Blot, RNA Binding Assay, Mouse Assay, Staining, Quantitative RT-PCR