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  • 99
    Thermo Fisher real time polymerase chain reaction qrt pcr
    Bleomycin induces collagen deposition in the lungs and reduces survival in Myr-Akt mice. Myr-Akt and wild type (WT) littermate mice were treated twice a week with bleomycin (0.035 U/g) or PBS (vehicle) for 4 weeks, as described in Material and Methods. (a) Mouse survival was recorded and plotted using a Kaplan-Meier curve. Myr-Akt mice had worse survival than WT mice in response to bleomycin treatment. N = 7 mice for each WT and Myr-Akt mouse group. (b) All surviving mice were sacrificed at day 33 and the lungs formalin-fixed, sectioned, and stained with Masson’s Trichrome. Five images per slide were captured. Red staining represents keratin, light red and pink staining represents cytoplasm, dark brown represents cell nuclei, and blue staining represents collagen. The images are representative images from 3 mice per group using a 20X objective (200× magnification). (c) The images were quantified blindly by histogram analysis using Adobe Photoshop CS and the percent of blue staining per high-power field (HPF) from ten images and expressed as mean ± SEM. (d) Lungs from WT and Myr-Akt mice subjected to PBS or bleomycin stained with H E, Trichrome, and F4/80 were evaluated by a board-certified pathologist for inflammation score (0 to 5 point scale). (e) Total RNA was extracted from homogenized lung tissue using TRIzol. cDNA was synthesized and <t>qRT-PCR</t> performed for mouse collagen IA and IIIB. Data are expressed as mean relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM and N = 4 mice per group.
    Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 950 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher transcript polymerase chain reaction qrt pcr kit
    Bleomycin induces collagen deposition in the lungs and reduces survival in Myr-Akt mice. Myr-Akt and wild type (WT) littermate mice were treated twice a week with bleomycin (0.035 U/g) or PBS (vehicle) for 4 weeks, as described in Material and Methods. (a) Mouse survival was recorded and plotted using a Kaplan-Meier curve. Myr-Akt mice had worse survival than WT mice in response to bleomycin treatment. N = 7 mice for each WT and Myr-Akt mouse group. (b) All surviving mice were sacrificed at day 33 and the lungs formalin-fixed, sectioned, and stained with Masson’s Trichrome. Five images per slide were captured. Red staining represents keratin, light red and pink staining represents cytoplasm, dark brown represents cell nuclei, and blue staining represents collagen. The images are representative images from 3 mice per group using a 20X objective (200× magnification). (c) The images were quantified blindly by histogram analysis using Adobe Photoshop CS and the percent of blue staining per high-power field (HPF) from ten images and expressed as mean ± SEM. (d) Lungs from WT and Myr-Akt mice subjected to PBS or bleomycin stained with H E, Trichrome, and F4/80 were evaluated by a board-certified pathologist for inflammation score (0 to 5 point scale). (e) Total RNA was extracted from homogenized lung tissue using TRIzol. cDNA was synthesized and <t>qRT-PCR</t> performed for mouse collagen IA and IIIB. Data are expressed as mean relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM and N = 4 mice per group.
    Transcript Polymerase Chain Reaction Qrt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 82/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa real time polymerase chain reaction qrt pcr
    Knockdown of FOXQ1 partially inhibited TGF-β1-induced aggressive tumor behaviors in SW480 cells and the changes in expression of downstream genes. ( A ) NC- and FOXQ1-shRNA cells were incubated in growth media supplemented with TGF-β1 (3 ng/ml) for 3 days. FOXQ1 expression was measured by <t>qRT-PCR</t> and Western blot. ( B ) The culture supernatant from NC-shRNA cells cultured for 3 days or NC- and FOXQ1-shRNA cells cultured for 5 days containing TGF-β1 (3 ng/ml) was collected and used in an in vitro angiogenesis assay. The vessel formation was measured in NC-shRNA-transduced EA.hy926 cells. The number of vessels was counted by fluorescent microscopy (120X). ( C ) FOXQ1- and NC-shRNA cells were treated with TGF-β1 (3 ng/ml) for 5 days, and cell morphology was examined by optical microscopy (120X) (upper panel). For the invasion assay, FOXQ1- and NC-shRNA cells were pre-treated with TGF-β1 (3 ng/ml) for 3 days before being seeded in the upper chamber of Transwell. After 36 hours, the cells migrating through the insert membrane were fixed and stained by crystal violet. The number of migrated cells was counted by optical microscopy (120X) (lower panels). ( D ) For the drug sensitivity assay, FOXQ1- and NC-shRNA cells were incubated in TGF-β1-containing media (3 ng/ml) for 4 days, and 5-FU or L-OHP (100 μg/ml) was added to the cell culture for an additional 24 hours. Flow cytometry was performed to measure cell apoptosis. ( E ) Expression of Wnt downstream target genes and EMT-associated markers in TGF-β1 treated NC- and FOXQ1-shRNA cells were measured by qRT-PCR and Western blot. The results are expressed as mean ± SD *P
    Real Time Polymerase Chain Reaction Qrt Pcr, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 223 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher reverse transcriptase polymerase chain reaction qrt pcr
    Knockdown of FOXQ1 partially inhibited TGF-β1-induced aggressive tumor behaviors in SW480 cells and the changes in expression of downstream genes. ( A ) NC- and FOXQ1-shRNA cells were incubated in growth media supplemented with TGF-β1 (3 ng/ml) for 3 days. FOXQ1 expression was measured by <t>qRT-PCR</t> and Western blot. ( B ) The culture supernatant from NC-shRNA cells cultured for 3 days or NC- and FOXQ1-shRNA cells cultured for 5 days containing TGF-β1 (3 ng/ml) was collected and used in an in vitro angiogenesis assay. The vessel formation was measured in NC-shRNA-transduced EA.hy926 cells. The number of vessels was counted by fluorescent microscopy (120X). ( C ) FOXQ1- and NC-shRNA cells were treated with TGF-β1 (3 ng/ml) for 5 days, and cell morphology was examined by optical microscopy (120X) (upper panel). For the invasion assay, FOXQ1- and NC-shRNA cells were pre-treated with TGF-β1 (3 ng/ml) for 3 days before being seeded in the upper chamber of Transwell. After 36 hours, the cells migrating through the insert membrane were fixed and stained by crystal violet. The number of migrated cells was counted by optical microscopy (120X) (lower panels). ( D ) For the drug sensitivity assay, FOXQ1- and NC-shRNA cells were incubated in TGF-β1-containing media (3 ng/ml) for 4 days, and 5-FU or L-OHP (100 μg/ml) was added to the cell culture for an additional 24 hours. Flow cytometry was performed to measure cell apoptosis. ( E ) Expression of Wnt downstream target genes and EMT-associated markers in TGF-β1 treated NC- and FOXQ1-shRNA cells were measured by qRT-PCR and Western blot. The results are expressed as mean ± SD *P
    Reverse Transcriptase Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 152 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad real time polymerase chain reaction qrt pcr
    Knockdown of FOXQ1 partially inhibited TGF-β1-induced aggressive tumor behaviors in SW480 cells and the changes in expression of downstream genes. ( A ) NC- and FOXQ1-shRNA cells were incubated in growth media supplemented with TGF-β1 (3 ng/ml) for 3 days. FOXQ1 expression was measured by <t>qRT-PCR</t> and Western blot. ( B ) The culture supernatant from NC-shRNA cells cultured for 3 days or NC- and FOXQ1-shRNA cells cultured for 5 days containing TGF-β1 (3 ng/ml) was collected and used in an in vitro angiogenesis assay. The vessel formation was measured in NC-shRNA-transduced EA.hy926 cells. The number of vessels was counted by fluorescent microscopy (120X). ( C ) FOXQ1- and NC-shRNA cells were treated with TGF-β1 (3 ng/ml) for 5 days, and cell morphology was examined by optical microscopy (120X) (upper panel). For the invasion assay, FOXQ1- and NC-shRNA cells were pre-treated with TGF-β1 (3 ng/ml) for 3 days before being seeded in the upper chamber of Transwell. After 36 hours, the cells migrating through the insert membrane were fixed and stained by crystal violet. The number of migrated cells was counted by optical microscopy (120X) (lower panels). ( D ) For the drug sensitivity assay, FOXQ1- and NC-shRNA cells were incubated in TGF-β1-containing media (3 ng/ml) for 4 days, and 5-FU or L-OHP (100 μg/ml) was added to the cell culture for an additional 24 hours. Flow cytometry was performed to measure cell apoptosis. ( E ) Expression of Wnt downstream target genes and EMT-associated markers in TGF-β1 treated NC- and FOXQ1-shRNA cells were measured by qRT-PCR and Western blot. The results are expressed as mean ± SD *P
    Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 228 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche real time polymerase chain reaction qrt pcr
    Knockdown of FOXQ1 partially inhibited TGF-β1-induced aggressive tumor behaviors in SW480 cells and the changes in expression of downstream genes. ( A ) NC- and FOXQ1-shRNA cells were incubated in growth media supplemented with TGF-β1 (3 ng/ml) for 3 days. FOXQ1 expression was measured by <t>qRT-PCR</t> and Western blot. ( B ) The culture supernatant from NC-shRNA cells cultured for 3 days or NC- and FOXQ1-shRNA cells cultured for 5 days containing TGF-β1 (3 ng/ml) was collected and used in an in vitro angiogenesis assay. The vessel formation was measured in NC-shRNA-transduced EA.hy926 cells. The number of vessels was counted by fluorescent microscopy (120X). ( C ) FOXQ1- and NC-shRNA cells were treated with TGF-β1 (3 ng/ml) for 5 days, and cell morphology was examined by optical microscopy (120X) (upper panel). For the invasion assay, FOXQ1- and NC-shRNA cells were pre-treated with TGF-β1 (3 ng/ml) for 3 days before being seeded in the upper chamber of Transwell. After 36 hours, the cells migrating through the insert membrane were fixed and stained by crystal violet. The number of migrated cells was counted by optical microscopy (120X) (lower panels). ( D ) For the drug sensitivity assay, FOXQ1- and NC-shRNA cells were incubated in TGF-β1-containing media (3 ng/ml) for 4 days, and 5-FU or L-OHP (100 μg/ml) was added to the cell culture for an additional 24 hours. Flow cytometry was performed to measure cell apoptosis. ( E ) Expression of Wnt downstream target genes and EMT-associated markers in TGF-β1 treated NC- and FOXQ1-shRNA cells were measured by qRT-PCR and Western blot. The results are expressed as mean ± SD *P
    Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 159 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad quantitative real time polymerase chain reaction qrt pcr
    Knockdown of FOXQ1 partially inhibited TGF-β1-induced aggressive tumor behaviors in SW480 cells and the changes in expression of downstream genes. ( A ) NC- and FOXQ1-shRNA cells were incubated in growth media supplemented with TGF-β1 (3 ng/ml) for 3 days. FOXQ1 expression was measured by <t>qRT-PCR</t> and Western blot. ( B ) The culture supernatant from NC-shRNA cells cultured for 3 days or NC- and FOXQ1-shRNA cells cultured for 5 days containing TGF-β1 (3 ng/ml) was collected and used in an in vitro angiogenesis assay. The vessel formation was measured in NC-shRNA-transduced EA.hy926 cells. The number of vessels was counted by fluorescent microscopy (120X). ( C ) FOXQ1- and NC-shRNA cells were treated with TGF-β1 (3 ng/ml) for 5 days, and cell morphology was examined by optical microscopy (120X) (upper panel). For the invasion assay, FOXQ1- and NC-shRNA cells were pre-treated with TGF-β1 (3 ng/ml) for 3 days before being seeded in the upper chamber of Transwell. After 36 hours, the cells migrating through the insert membrane were fixed and stained by crystal violet. The number of migrated cells was counted by optical microscopy (120X) (lower panels). ( D ) For the drug sensitivity assay, FOXQ1- and NC-shRNA cells were incubated in TGF-β1-containing media (3 ng/ml) for 4 days, and 5-FU or L-OHP (100 μg/ml) was added to the cell culture for an additional 24 hours. Flow cytometry was performed to measure cell apoptosis. ( E ) Expression of Wnt downstream target genes and EMT-associated markers in TGF-β1 treated NC- and FOXQ1-shRNA cells were measured by qRT-PCR and Western blot. The results are expressed as mean ± SD *P
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 520 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies quantitative real time polymerase chain reaction qrt pcr
    Knockdown of FOXQ1 partially inhibited TGF-β1-induced aggressive tumor behaviors in SW480 cells and the changes in expression of downstream genes. ( A ) NC- and FOXQ1-shRNA cells were incubated in growth media supplemented with TGF-β1 (3 ng/ml) for 3 days. FOXQ1 expression was measured by <t>qRT-PCR</t> and Western blot. ( B ) The culture supernatant from NC-shRNA cells cultured for 3 days or NC- and FOXQ1-shRNA cells cultured for 5 days containing TGF-β1 (3 ng/ml) was collected and used in an in vitro angiogenesis assay. The vessel formation was measured in NC-shRNA-transduced EA.hy926 cells. The number of vessels was counted by fluorescent microscopy (120X). ( C ) FOXQ1- and NC-shRNA cells were treated with TGF-β1 (3 ng/ml) for 5 days, and cell morphology was examined by optical microscopy (120X) (upper panel). For the invasion assay, FOXQ1- and NC-shRNA cells were pre-treated with TGF-β1 (3 ng/ml) for 3 days before being seeded in the upper chamber of Transwell. After 36 hours, the cells migrating through the insert membrane were fixed and stained by crystal violet. The number of migrated cells was counted by optical microscopy (120X) (lower panels). ( D ) For the drug sensitivity assay, FOXQ1- and NC-shRNA cells were incubated in TGF-β1-containing media (3 ng/ml) for 4 days, and 5-FU or L-OHP (100 μg/ml) was added to the cell culture for an additional 24 hours. Flow cytometry was performed to measure cell apoptosis. ( E ) Expression of Wnt downstream target genes and EMT-associated markers in TGF-β1 treated NC- and FOXQ1-shRNA cells were measured by qRT-PCR and Western blot. The results are expressed as mean ± SD *P
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Agilent technologies, 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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    Thermo Fisher reverse transcription polymerase chain reaction qrt pcr module
    Knockdown of FOXQ1 partially inhibited TGF-β1-induced aggressive tumor behaviors in SW480 cells and the changes in expression of downstream genes. ( A ) NC- and FOXQ1-shRNA cells were incubated in growth media supplemented with TGF-β1 (3 ng/ml) for 3 days. FOXQ1 expression was measured by <t>qRT-PCR</t> and Western blot. ( B ) The culture supernatant from NC-shRNA cells cultured for 3 days or NC- and FOXQ1-shRNA cells cultured for 5 days containing TGF-β1 (3 ng/ml) was collected and used in an in vitro angiogenesis assay. The vessel formation was measured in NC-shRNA-transduced EA.hy926 cells. The number of vessels was counted by fluorescent microscopy (120X). ( C ) FOXQ1- and NC-shRNA cells were treated with TGF-β1 (3 ng/ml) for 5 days, and cell morphology was examined by optical microscopy (120X) (upper panel). For the invasion assay, FOXQ1- and NC-shRNA cells were pre-treated with TGF-β1 (3 ng/ml) for 3 days before being seeded in the upper chamber of Transwell. After 36 hours, the cells migrating through the insert membrane were fixed and stained by crystal violet. The number of migrated cells was counted by optical microscopy (120X) (lower panels). ( D ) For the drug sensitivity assay, FOXQ1- and NC-shRNA cells were incubated in TGF-β1-containing media (3 ng/ml) for 4 days, and 5-FU or L-OHP (100 μg/ml) was added to the cell culture for an additional 24 hours. Flow cytometry was performed to measure cell apoptosis. ( E ) Expression of Wnt downstream target genes and EMT-associated markers in TGF-β1 treated NC- and FOXQ1-shRNA cells were measured by qRT-PCR and Western blot. The results are expressed as mean ± SD *P
    Reverse Transcription Polymerase Chain Reaction Qrt Pcr Module, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher quantitative reverse transcription polymerase chain reaction qrt pcr
    Knockdown of FOXQ1 partially inhibited TGF-β1-induced aggressive tumor behaviors in SW480 cells and the changes in expression of downstream genes. ( A ) NC- and FOXQ1-shRNA cells were incubated in growth media supplemented with TGF-β1 (3 ng/ml) for 3 days. FOXQ1 expression was measured by <t>qRT-PCR</t> and Western blot. ( B ) The culture supernatant from NC-shRNA cells cultured for 3 days or NC- and FOXQ1-shRNA cells cultured for 5 days containing TGF-β1 (3 ng/ml) was collected and used in an in vitro angiogenesis assay. The vessel formation was measured in NC-shRNA-transduced EA.hy926 cells. The number of vessels was counted by fluorescent microscopy (120X). ( C ) FOXQ1- and NC-shRNA cells were treated with TGF-β1 (3 ng/ml) for 5 days, and cell morphology was examined by optical microscopy (120X) (upper panel). For the invasion assay, FOXQ1- and NC-shRNA cells were pre-treated with TGF-β1 (3 ng/ml) for 3 days before being seeded in the upper chamber of Transwell. After 36 hours, the cells migrating through the insert membrane were fixed and stained by crystal violet. The number of migrated cells was counted by optical microscopy (120X) (lower panels). ( D ) For the drug sensitivity assay, FOXQ1- and NC-shRNA cells were incubated in TGF-β1-containing media (3 ng/ml) for 4 days, and 5-FU or L-OHP (100 μg/ml) was added to the cell culture for an additional 24 hours. Flow cytometry was performed to measure cell apoptosis. ( E ) Expression of Wnt downstream target genes and EMT-associated markers in TGF-β1 treated NC- and FOXQ1-shRNA cells were measured by qRT-PCR and Western blot. The results are expressed as mean ± SD *P
    Quantitative Reverse Transcription Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1237 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Genewiz reverse transcription polymerase chain reaction qrt pcr validation
    FA2H expression among breast cancer cell lines. (A) FA2H gene expression among different breast cancer cell lines by <t>qRT-PCR.</t> (B) FA2H protein expression among different breast cancer cell lines by Western-blotting. (C) FA2H protein expression quantified using the ImageJ software. (D) FA2H gene expression across 56 breast cancer cell lines ( 26 ).
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    Thermo Fisher quantificational reverse transcription polymerase chain reaction qrt pcr trizol
    FA2H expression among breast cancer cell lines. (A) FA2H gene expression among different breast cancer cell lines by <t>qRT-PCR.</t> (B) FA2H protein expression among different breast cancer cell lines by Western-blotting. (C) FA2H protein expression quantified using the ImageJ software. (D) FA2H gene expression across 56 breast cancer cell lines ( 26 ).
    Quantificational Reverse Transcription Polymerase Chain Reaction Qrt Pcr Trizol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 80/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad real time polymerase chain reaction qrt pcr detection system
    FA2H expression among breast cancer cell lines. (A) FA2H gene expression among different breast cancer cell lines by <t>qRT-PCR.</t> (B) FA2H protein expression among different breast cancer cell lines by Western-blotting. (C) FA2H protein expression quantified using the ImageJ software. (D) FA2H gene expression across 56 breast cancer cell lines ( 26 ).
    Real Time Polymerase Chain Reaction Qrt Pcr Detection System, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 82/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher quantitative real time polymerase chain reaction qrt pcr trizol
    FA2H expression among breast cancer cell lines. (A) FA2H gene expression among different breast cancer cell lines by <t>qRT-PCR.</t> (B) FA2H protein expression among different breast cancer cell lines by Western-blotting. (C) FA2H protein expression quantified using the ImageJ software. (D) FA2H gene expression across 56 breast cancer cell lines ( 26 ).
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr Trizol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 80/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa quantitative real time polymerase chain reaction qrt pcr kits
    Detection of the expression of miR-218 and miR-34a in tissue specimens by <t>qRT-PCR.</t> Compared with those in normal cervical tissues, the expression levels of miR-34a and miR-218 in cervical cancer tissues are significantly decreased, **P
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr Kits, supplied by TaKaRa, used in various techniques. Bioz Stars score: 82/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa real time polymerase chain reaction qrt pcr reagents
    Detection of the expression of miR-218 and miR-34a in tissue specimens by <t>qRT-PCR.</t> Compared with those in normal cervical tissues, the expression levels of miR-34a and miR-218 in cervical cancer tissues are significantly decreased, **P
    Real Time Polymerase Chain Reaction Qrt Pcr Reagents, supplied by TaKaRa, used in various techniques. Bioz Stars score: 82/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher real time polymerase chain reaction qrt pcr reagents
    Detection of the expression of miR-218 and miR-34a in tissue specimens by <t>qRT-PCR.</t> Compared with those in normal cervical tissues, the expression levels of miR-34a and miR-218 in cervical cancer tissues are significantly decreased, **P
    Real Time Polymerase Chain Reaction Qrt Pcr Reagents, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 82/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher real time quantitative polymerase chain reaction qrt pcr
    Detection of the expression of miR-218 and miR-34a in tissue specimens by <t>qRT-PCR.</t> Compared with those in normal cervical tissues, the expression levels of miR-34a and miR-218 in cervical cancer tissues are significantly decreased, **P
    Real Time Quantitative Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 124 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher quantitative real time polymerase chain reaction qrt pcr trizol reagent
    Detection of the expression of miR-218 and miR-34a in tissue specimens by <t>qRT-PCR.</t> Compared with those in normal cervical tissues, the expression levels of miR-34a and miR-218 in cervical cancer tissues are significantly decreased, **P
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr Trizol Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Bleomycin induces collagen deposition in the lungs and reduces survival in Myr-Akt mice. Myr-Akt and wild type (WT) littermate mice were treated twice a week with bleomycin (0.035 U/g) or PBS (vehicle) for 4 weeks, as described in Material and Methods. (a) Mouse survival was recorded and plotted using a Kaplan-Meier curve. Myr-Akt mice had worse survival than WT mice in response to bleomycin treatment. N = 7 mice for each WT and Myr-Akt mouse group. (b) All surviving mice were sacrificed at day 33 and the lungs formalin-fixed, sectioned, and stained with Masson’s Trichrome. Five images per slide were captured. Red staining represents keratin, light red and pink staining represents cytoplasm, dark brown represents cell nuclei, and blue staining represents collagen. The images are representative images from 3 mice per group using a 20X objective (200× magnification). (c) The images were quantified blindly by histogram analysis using Adobe Photoshop CS and the percent of blue staining per high-power field (HPF) from ten images and expressed as mean ± SEM. (d) Lungs from WT and Myr-Akt mice subjected to PBS or bleomycin stained with H E, Trichrome, and F4/80 were evaluated by a board-certified pathologist for inflammation score (0 to 5 point scale). (e) Total RNA was extracted from homogenized lung tissue using TRIzol. cDNA was synthesized and qRT-PCR performed for mouse collagen IA and IIIB. Data are expressed as mean relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM and N = 4 mice per group.

    Journal: Advances in bioscience and biotechnology (Print)

    Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy

    doi: 10.4236/abb.2019.1010027

    Figure Lengend Snippet: Bleomycin induces collagen deposition in the lungs and reduces survival in Myr-Akt mice. Myr-Akt and wild type (WT) littermate mice were treated twice a week with bleomycin (0.035 U/g) or PBS (vehicle) for 4 weeks, as described in Material and Methods. (a) Mouse survival was recorded and plotted using a Kaplan-Meier curve. Myr-Akt mice had worse survival than WT mice in response to bleomycin treatment. N = 7 mice for each WT and Myr-Akt mouse group. (b) All surviving mice were sacrificed at day 33 and the lungs formalin-fixed, sectioned, and stained with Masson’s Trichrome. Five images per slide were captured. Red staining represents keratin, light red and pink staining represents cytoplasm, dark brown represents cell nuclei, and blue staining represents collagen. The images are representative images from 3 mice per group using a 20X objective (200× magnification). (c) The images were quantified blindly by histogram analysis using Adobe Photoshop CS and the percent of blue staining per high-power field (HPF) from ten images and expressed as mean ± SEM. (d) Lungs from WT and Myr-Akt mice subjected to PBS or bleomycin stained with H E, Trichrome, and F4/80 were evaluated by a board-certified pathologist for inflammation score (0 to 5 point scale). (e) Total RNA was extracted from homogenized lung tissue using TRIzol. cDNA was synthesized and qRT-PCR performed for mouse collagen IA and IIIB. Data are expressed as mean relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM and N = 4 mice per group.

    Article Snippet: Primers and SYBR Green Master Mix (4385610) for quantitative real-time polymerase chain reaction (qRT-PCR) were purchased from Thermofisher Scientific unless indicated otherwise.

    Techniques: Mouse Assay, Staining, Synthesized, Quantitative RT-PCR, IA, Expressing

    Reduced autophagy in the lungs of Myr-Akt mice. (a) 6 – 8 week old Myr-Akt or WT mice were sacrificed and lungs harvested, paraffin embedded, and subjected to LC3B immunohistochemical staining. Ten pictures per section were imaged using 10X and 40X objectives (100× and 400× magnification, respectively). (b) Images were quantified using histogram analysis feature in Adobe Photoshop CS5 software for percent brown stain (F4/80+ pixels) per high-power field (HPF) from at least five images per section and expressed as mean ± SEM. (c) Total RNA was extracted from homogenized lungs using TRIzol. cDNA was synthesized and qRT-PCR performed using primers specific for mouse Beclin-1 , Lc 3 a , and Lc 3 b mRNAs. Data are expressed as relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM. (d) Total protein were isolated and equal amounts of protein separated on a 4% – 12% SDS-PAGE gel and immunoblotted for BECLIN-1, LC3A/B-I and LC3A/B-II, and β -ACTIN. (e) Total RNA was extracted from homogenized lungs using TRIzol. cDNA was synthesized and qRT-PCR performed using primers specific for mouse p 62/ SQSTM1 mRNA. Data are expressed as relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM. (f) Immunoblot for phosphor-mTOR (ser2448) and mTOR (Cell Signaling Technology). Bands were quantified using densitometry by comparing band density over β -ACTIN or total mTOR in each respective lane. Data are expressed as the mean densitometry value ± SEM. 4 pairs of age-matched littermate mice were used for western analysis and 5 pairs of age-matched littermate mice were used for gene expression.

    Journal: Advances in bioscience and biotechnology (Print)

    Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy

    doi: 10.4236/abb.2019.1010027

    Figure Lengend Snippet: Reduced autophagy in the lungs of Myr-Akt mice. (a) 6 – 8 week old Myr-Akt or WT mice were sacrificed and lungs harvested, paraffin embedded, and subjected to LC3B immunohistochemical staining. Ten pictures per section were imaged using 10X and 40X objectives (100× and 400× magnification, respectively). (b) Images were quantified using histogram analysis feature in Adobe Photoshop CS5 software for percent brown stain (F4/80+ pixels) per high-power field (HPF) from at least five images per section and expressed as mean ± SEM. (c) Total RNA was extracted from homogenized lungs using TRIzol. cDNA was synthesized and qRT-PCR performed using primers specific for mouse Beclin-1 , Lc 3 a , and Lc 3 b mRNAs. Data are expressed as relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM. (d) Total protein were isolated and equal amounts of protein separated on a 4% – 12% SDS-PAGE gel and immunoblotted for BECLIN-1, LC3A/B-I and LC3A/B-II, and β -ACTIN. (e) Total RNA was extracted from homogenized lungs using TRIzol. cDNA was synthesized and qRT-PCR performed using primers specific for mouse p 62/ SQSTM1 mRNA. Data are expressed as relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM. (f) Immunoblot for phosphor-mTOR (ser2448) and mTOR (Cell Signaling Technology). Bands were quantified using densitometry by comparing band density over β -ACTIN or total mTOR in each respective lane. Data are expressed as the mean densitometry value ± SEM. 4 pairs of age-matched littermate mice were used for western analysis and 5 pairs of age-matched littermate mice were used for gene expression.

    Article Snippet: Primers and SYBR Green Master Mix (4385610) for quantitative real-time polymerase chain reaction (qRT-PCR) were purchased from Thermofisher Scientific unless indicated otherwise.

    Techniques: Mouse Assay, Immunohistochemistry, Staining, Software, Synthesized, Quantitative RT-PCR, Expressing, Isolation, SDS Page, Western Blot

    Differential myofibroblast expression in the lungs of Myr-Akt and WT mice. (a) 6 – 8 week old Myr-Akt or WT mice were sacrificed and lungs harvested, paraffin embedded, and subjected to α -SMA immunohistochemical staining for myofibroblasts. Ten pictures per section were imaged using 4X and 20X objectives (40× and 200× magnification, respectively). (b) Images were quantified by histogram analysis using Adobe Photoshop CS5 software and percent brown stain ( α -SMA+ pixels) per high-power field (HPF) from at least five images per section are expressed as mean ± SEM. (c) Total RNA was extracted from Myr-Akt or WT littermate mouse lungs and α -SMA mRNA was measured by qRT-PCR. Data are expressed as relative copy number (RCN) of α -SMA mRNA expression over the average of endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represent the mean RCN ± SEM. Single-cell suspension from lung homogenate were immunostained using mouse α-SMA antibodies or double-immunostained with α -SMA and M-CSF-R (CD115) antibodies and analyzed by flow cytometry. (d) Data represents mean percent of α -SMA+ cells and (e) α -SMA+/M-CSF-R+ cells ± SEM. 5 pairs of age-matched littermate mice were used for each experiment.

    Journal: Advances in bioscience and biotechnology (Print)

    Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy

    doi: 10.4236/abb.2019.1010027

    Figure Lengend Snippet: Differential myofibroblast expression in the lungs of Myr-Akt and WT mice. (a) 6 – 8 week old Myr-Akt or WT mice were sacrificed and lungs harvested, paraffin embedded, and subjected to α -SMA immunohistochemical staining for myofibroblasts. Ten pictures per section were imaged using 4X and 20X objectives (40× and 200× magnification, respectively). (b) Images were quantified by histogram analysis using Adobe Photoshop CS5 software and percent brown stain ( α -SMA+ pixels) per high-power field (HPF) from at least five images per section are expressed as mean ± SEM. (c) Total RNA was extracted from Myr-Akt or WT littermate mouse lungs and α -SMA mRNA was measured by qRT-PCR. Data are expressed as relative copy number (RCN) of α -SMA mRNA expression over the average of endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represent the mean RCN ± SEM. Single-cell suspension from lung homogenate were immunostained using mouse α-SMA antibodies or double-immunostained with α -SMA and M-CSF-R (CD115) antibodies and analyzed by flow cytometry. (d) Data represents mean percent of α -SMA+ cells and (e) α -SMA+/M-CSF-R+ cells ± SEM. 5 pairs of age-matched littermate mice were used for each experiment.

    Article Snippet: Primers and SYBR Green Master Mix (4385610) for quantitative real-time polymerase chain reaction (qRT-PCR) were purchased from Thermofisher Scientific unless indicated otherwise.

    Techniques: Expressing, Mouse Assay, Immunohistochemistry, Staining, Software, Quantitative RT-PCR, Flow Cytometry, Cytometry

    Bleomycin suppresses autophagy gene expression in the lungs of Myr-Akt mice. Total RNA was extracted from paraffin embedded lung tissue section with RecoverAll Total Nucleic Acid Isolation Kit for FFPE according to manufacturer’s protocol. cDNA was synthesized and qRT-PCR performed for mouse Beclin- 1, Lc 3 a , and Lc 3 b . Data are expressed as relative copy number (RCN) of mRNA expression over average of endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh .). Data represents the mean RCN ± SEM and N = 4 mice per group.

    Journal: Advances in bioscience and biotechnology (Print)

    Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy

    doi: 10.4236/abb.2019.1010027

    Figure Lengend Snippet: Bleomycin suppresses autophagy gene expression in the lungs of Myr-Akt mice. Total RNA was extracted from paraffin embedded lung tissue section with RecoverAll Total Nucleic Acid Isolation Kit for FFPE according to manufacturer’s protocol. cDNA was synthesized and qRT-PCR performed for mouse Beclin- 1, Lc 3 a , and Lc 3 b . Data are expressed as relative copy number (RCN) of mRNA expression over average of endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh .). Data represents the mean RCN ± SEM and N = 4 mice per group.

    Article Snippet: Primers and SYBR Green Master Mix (4385610) for quantitative real-time polymerase chain reaction (qRT-PCR) were purchased from Thermofisher Scientific unless indicated otherwise.

    Techniques: Expressing, Mouse Assay, Isolation, Formalin-fixed Paraffin-Embedded, Synthesized, Quantitative RT-PCR

    Knockdown of FOXQ1 partially inhibited TGF-β1-induced aggressive tumor behaviors in SW480 cells and the changes in expression of downstream genes. ( A ) NC- and FOXQ1-shRNA cells were incubated in growth media supplemented with TGF-β1 (3 ng/ml) for 3 days. FOXQ1 expression was measured by qRT-PCR and Western blot. ( B ) The culture supernatant from NC-shRNA cells cultured for 3 days or NC- and FOXQ1-shRNA cells cultured for 5 days containing TGF-β1 (3 ng/ml) was collected and used in an in vitro angiogenesis assay. The vessel formation was measured in NC-shRNA-transduced EA.hy926 cells. The number of vessels was counted by fluorescent microscopy (120X). ( C ) FOXQ1- and NC-shRNA cells were treated with TGF-β1 (3 ng/ml) for 5 days, and cell morphology was examined by optical microscopy (120X) (upper panel). For the invasion assay, FOXQ1- and NC-shRNA cells were pre-treated with TGF-β1 (3 ng/ml) for 3 days before being seeded in the upper chamber of Transwell. After 36 hours, the cells migrating through the insert membrane were fixed and stained by crystal violet. The number of migrated cells was counted by optical microscopy (120X) (lower panels). ( D ) For the drug sensitivity assay, FOXQ1- and NC-shRNA cells were incubated in TGF-β1-containing media (3 ng/ml) for 4 days, and 5-FU or L-OHP (100 μg/ml) was added to the cell culture for an additional 24 hours. Flow cytometry was performed to measure cell apoptosis. ( E ) Expression of Wnt downstream target genes and EMT-associated markers in TGF-β1 treated NC- and FOXQ1-shRNA cells were measured by qRT-PCR and Western blot. The results are expressed as mean ± SD *P

    Journal: Cancer Biology & Therapy

    Article Title: FOXQ1 mediates the crosstalk between TGF-β and Wnt signaling pathways in the progression of colorectal cancer

    doi: 10.1080/15384047.2015.1047568

    Figure Lengend Snippet: Knockdown of FOXQ1 partially inhibited TGF-β1-induced aggressive tumor behaviors in SW480 cells and the changes in expression of downstream genes. ( A ) NC- and FOXQ1-shRNA cells were incubated in growth media supplemented with TGF-β1 (3 ng/ml) for 3 days. FOXQ1 expression was measured by qRT-PCR and Western blot. ( B ) The culture supernatant from NC-shRNA cells cultured for 3 days or NC- and FOXQ1-shRNA cells cultured for 5 days containing TGF-β1 (3 ng/ml) was collected and used in an in vitro angiogenesis assay. The vessel formation was measured in NC-shRNA-transduced EA.hy926 cells. The number of vessels was counted by fluorescent microscopy (120X). ( C ) FOXQ1- and NC-shRNA cells were treated with TGF-β1 (3 ng/ml) for 5 days, and cell morphology was examined by optical microscopy (120X) (upper panel). For the invasion assay, FOXQ1- and NC-shRNA cells were pre-treated with TGF-β1 (3 ng/ml) for 3 days before being seeded in the upper chamber of Transwell. After 36 hours, the cells migrating through the insert membrane were fixed and stained by crystal violet. The number of migrated cells was counted by optical microscopy (120X) (lower panels). ( D ) For the drug sensitivity assay, FOXQ1- and NC-shRNA cells were incubated in TGF-β1-containing media (3 ng/ml) for 4 days, and 5-FU or L-OHP (100 μg/ml) was added to the cell culture for an additional 24 hours. Flow cytometry was performed to measure cell apoptosis. ( E ) Expression of Wnt downstream target genes and EMT-associated markers in TGF-β1 treated NC- and FOXQ1-shRNA cells were measured by qRT-PCR and Western blot. The results are expressed as mean ± SD *P

    Article Snippet: All reagents and primers used in quantitative real-time polymerase chain reaction (qRT-PCR) were purchased from Takara Bio (Dalian, China).

    Techniques: Expressing, shRNA, Incubation, Quantitative RT-PCR, Western Blot, Cell Culture, In Vitro, Angiogenesis Assay, Microscopy, Invasion Assay, Staining, Sensitive Assay, Flow Cytometry, Cytometry

    FA2H expression among breast cancer cell lines. (A) FA2H gene expression among different breast cancer cell lines by qRT-PCR. (B) FA2H protein expression among different breast cancer cell lines by Western-blotting. (C) FA2H protein expression quantified using the ImageJ software. (D) FA2H gene expression across 56 breast cancer cell lines ( 26 ).

    Journal: Frontiers in Oncology

    Article Title: FA2H Exhibits Tumor Suppressive Roles on Breast Cancers via Cancer Stemness Control

    doi: 10.3389/fonc.2019.01089

    Figure Lengend Snippet: FA2H expression among breast cancer cell lines. (A) FA2H gene expression among different breast cancer cell lines by qRT-PCR. (B) FA2H protein expression among different breast cancer cell lines by Western-blotting. (C) FA2H protein expression quantified using the ImageJ software. (D) FA2H gene expression across 56 breast cancer cell lines ( 26 ).

    Article Snippet: Primers designed for candidate genes and used for quantitative reverse transcription polymerase chain reaction (qRT-PCR) validation (ordered from GENEWIZ) were listed in .

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Software

    Detection of the expression of miR-218 and miR-34a in tissue specimens by qRT-PCR. Compared with those in normal cervical tissues, the expression levels of miR-34a and miR-218 in cervical cancer tissues are significantly decreased, **P

    Journal: Oncology Letters

    Article Title: Values of miR-34a and miR-218 expression in the diagnosis of cervical cancer and the prediction of prognosis

    doi: 10.3892/ol.2018.7791

    Figure Lengend Snippet: Detection of the expression of miR-218 and miR-34a in tissue specimens by qRT-PCR. Compared with those in normal cervical tissues, the expression levels of miR-34a and miR-218 in cervical cancer tissues are significantly decreased, **P

    Article Snippet: The present study was approved by the Clinical Ethics Committee, and all the patients or family members signed the informed consent; TRIzol kit (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA); primer synthesis, reverse transcription kits and quantitative real-time polymerase chain reaction (qRT-PCR) kits (all from Takara Biotechnology Co., Ltd., Dalian, China).

    Techniques: Expressing, Quantitative RT-PCR

    Expression of NF-κB1 and Bcl-2 detection in glioma tissues by qRT-PCR. Notes: The expression of NF-κB1 in glioma was significantly higher than that in nonneoplastic brain tissues ( P

    Journal: OncoTargets and therapy

    Article Title: Nuclear factor-kappa B1 inhibits early apoptosis of glioma cells by promoting the expression of Bcl-2

    doi: 10.2147/OTT.S144014

    Figure Lengend Snippet: Expression of NF-κB1 and Bcl-2 detection in glioma tissues by qRT-PCR. Notes: The expression of NF-κB1 in glioma was significantly higher than that in nonneoplastic brain tissues ( P

    Article Snippet: Relative levels of NF-κB1 and Bcl-2 mRNA were examined using SYBR green quantitative real-time polymerase chain reaction (qRT-PCR) (LightCycler® 480; Hoffman-La Roche Ltd., Basel, Switzerland) and were normalized to levels of GAPDH mRNA.

    Techniques: Expressing, Quantitative RT-PCR

    HFD feeding increased the translocation to the nucleus of p65 and expression of NFκB‐related genes in Irf3 −/− mice. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expressions of phospho‐IKKα∕β, IKKβ, phospho‐p65, p65, and IκBα were assessed by western blot and normalized to GAPDH, n = 4 per genotype. (B) Formalin‐fixed paraffin‐embedded sections of liver were deparaffinized, and localization of phospho‐p65 was assessed by immunohistochemistry. Images were acquired at 40×. White arrows indicate nonparenchymal cells, and black arrows indicate hepatocytes. Images are representative of triplicate images from eight mice per treatment group. Values represent means ± SEM. (C) Nuclear fractions were isolated from livers, and equal amounts of protein were assessed by western blot and probed against phosphor‐Ser 536 p65‐NFκB. Lamin A/C was used as a nuclei marker, and GAPDH was used as a cytosolic marker. (D) Expressions of CCL5, CXCL10, CXCL2, and CCL3 were analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values with different superscripts are significantly different from each other, P

    Journal: Hepatology Communications

    Article Title: Nontranscriptional Activity of Interferon Regulatory Factor 3 Protects Mice From High‐Fat Diet‐Induced Liver Injury

    doi: 10.1002/hep4.1441

    Figure Lengend Snippet: HFD feeding increased the translocation to the nucleus of p65 and expression of NFκB‐related genes in Irf3 −/− mice. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expressions of phospho‐IKKα∕β, IKKβ, phospho‐p65, p65, and IκBα were assessed by western blot and normalized to GAPDH, n = 4 per genotype. (B) Formalin‐fixed paraffin‐embedded sections of liver were deparaffinized, and localization of phospho‐p65 was assessed by immunohistochemistry. Images were acquired at 40×. White arrows indicate nonparenchymal cells, and black arrows indicate hepatocytes. Images are representative of triplicate images from eight mice per treatment group. Values represent means ± SEM. (C) Nuclear fractions were isolated from livers, and equal amounts of protein were assessed by western blot and probed against phosphor‐Ser 536 p65‐NFκB. Lamin A/C was used as a nuclei marker, and GAPDH was used as a cytosolic marker. (D) Expressions of CCL5, CXCL10, CXCL2, and CCL3 were analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values with different superscripts are significantly different from each other, P

    Article Snippet: Two micrograms of liver RNA was reverse transcribed and analyzed with PowerSYBR quantitative real‐time polymerase chain reaction (qRT‐PCR) kits (Applied Biosystems, Foster City, CA) on a QuantStudio5 RT‐PCR Machine (Applied Biosystems).

    Techniques: Translocation Assay, Expressing, Mouse Assay, Western Blot, Formalin-fixed Paraffin-Embedded, Immunohistochemistry, Isolation, Marker, Quantitative RT-PCR

    Irf3 S1/S1 mice reduced HFD‐induced liver injury, steatosis, and markers of inflammation. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A,B) ALT and AST activities were measured in plasma, n = 12 per genotype. (C) Hepatic triglyceride content was measured in whole‐liver homogenates, n = 12 per genotype. (D) Paraffin‐embedded liver sections were stained with hematoxylin and eosin. Images were acquired using 10× and 40× objectives, n = 4 per genotype. Expressions of (E) TNFα, (F) IL‐1β, and (G) MCP‐1 mRNA were analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Journal: Hepatology Communications

    Article Title: Nontranscriptional Activity of Interferon Regulatory Factor 3 Protects Mice From High‐Fat Diet‐Induced Liver Injury

    doi: 10.1002/hep4.1441

    Figure Lengend Snippet: Irf3 S1/S1 mice reduced HFD‐induced liver injury, steatosis, and markers of inflammation. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A,B) ALT and AST activities were measured in plasma, n = 12 per genotype. (C) Hepatic triglyceride content was measured in whole‐liver homogenates, n = 12 per genotype. (D) Paraffin‐embedded liver sections were stained with hematoxylin and eosin. Images were acquired using 10× and 40× objectives, n = 4 per genotype. Expressions of (E) TNFα, (F) IL‐1β, and (G) MCP‐1 mRNA were analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Article Snippet: Two micrograms of liver RNA was reverse transcribed and analyzed with PowerSYBR quantitative real‐time polymerase chain reaction (qRT‐PCR) kits (Applied Biosystems, Foster City, CA) on a QuantStudio5 RT‐PCR Machine (Applied Biosystems).

    Techniques: Mouse Assay, AST Assay, Staining, Quantitative RT-PCR

    HFD enhanced CD45 + infiltration in Irf3 −/− but not in wild‐type and Irf3 S1/S1 livers. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expression of CD45 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 8 per genotype. (B) Hepatic CD45 + leukocytes were gated in isolated liver NPCs by flow cytometry, and total number of cells were quantified in isolated liver NPCs, n = 4 per genotype. (C) Paraffin‐embedded liver sections were stained for CD45. Images were acquired using a 20× objective, n = 4 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Journal: Hepatology Communications

    Article Title: Nontranscriptional Activity of Interferon Regulatory Factor 3 Protects Mice From High‐Fat Diet‐Induced Liver Injury

    doi: 10.1002/hep4.1441

    Figure Lengend Snippet: HFD enhanced CD45 + infiltration in Irf3 −/− but not in wild‐type and Irf3 S1/S1 livers. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expression of CD45 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 8 per genotype. (B) Hepatic CD45 + leukocytes were gated in isolated liver NPCs by flow cytometry, and total number of cells were quantified in isolated liver NPCs, n = 4 per genotype. (C) Paraffin‐embedded liver sections were stained for CD45. Images were acquired using a 20× objective, n = 4 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Article Snippet: Two micrograms of liver RNA was reverse transcribed and analyzed with PowerSYBR quantitative real‐time polymerase chain reaction (qRT‐PCR) kits (Applied Biosystems, Foster City, CA) on a QuantStudio5 RT‐PCR Machine (Applied Biosystems).

    Techniques: Mouse Assay, Expressing, Quantitative RT-PCR, Isolation, Flow Cytometry, Cytometry, Staining

    Enhanced markers of hepatocellular death and fibrosis in Irf3 −/− mice compared with wild‐type and Irf3 S1/S1 mice after HFD feeding. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A,B) Paraffin‐embedded liver sections were stained for M30, a caspase‐dependent cleavage product of cytokeratin 18. Images were acquired using 10× and 40× objectives, and the positive area was quantified, n = 4 per genotype. (C,D) Paraffin‐embedded liver sections were stained with sirius red. Images were acquired using a 40× objective, and the positive area was quantified, n = 4 per genotype. (E) Expression of Col1α1 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 8 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Journal: Hepatology Communications

    Article Title: Nontranscriptional Activity of Interferon Regulatory Factor 3 Protects Mice From High‐Fat Diet‐Induced Liver Injury

    doi: 10.1002/hep4.1441

    Figure Lengend Snippet: Enhanced markers of hepatocellular death and fibrosis in Irf3 −/− mice compared with wild‐type and Irf3 S1/S1 mice after HFD feeding. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A,B) Paraffin‐embedded liver sections were stained for M30, a caspase‐dependent cleavage product of cytokeratin 18. Images were acquired using 10× and 40× objectives, and the positive area was quantified, n = 4 per genotype. (C,D) Paraffin‐embedded liver sections were stained with sirius red. Images were acquired using a 40× objective, and the positive area was quantified, n = 4 per genotype. (E) Expression of Col1α1 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 8 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Article Snippet: Two micrograms of liver RNA was reverse transcribed and analyzed with PowerSYBR quantitative real‐time polymerase chain reaction (qRT‐PCR) kits (Applied Biosystems, Foster City, CA) on a QuantStudio5 RT‐PCR Machine (Applied Biosystems).

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

    Hepatic IRF3 activation after HFD feeding. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expression of IFIT1 and IFIT3 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. (B) Expression of IRF3 was assessed by western blot and normalized to GAPDH, n = 4 per genotype. (C) Expression of IRF3 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Journal: Hepatology Communications

    Article Title: Nontranscriptional Activity of Interferon Regulatory Factor 3 Protects Mice From High‐Fat Diet‐Induced Liver Injury

    doi: 10.1002/hep4.1441

    Figure Lengend Snippet: Hepatic IRF3 activation after HFD feeding. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expression of IFIT1 and IFIT3 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. (B) Expression of IRF3 was assessed by western blot and normalized to GAPDH, n = 4 per genotype. (C) Expression of IRF3 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Article Snippet: Two micrograms of liver RNA was reverse transcribed and analyzed with PowerSYBR quantitative real‐time polymerase chain reaction (qRT‐PCR) kits (Applied Biosystems, Foster City, CA) on a QuantStudio5 RT‐PCR Machine (Applied Biosystems).

    Techniques: Activation Assay, Mouse Assay, Expressing, Quantitative RT-PCR, Western Blot

    ncU87-Exo and shU87-Exo are internalized by HUVECs. (A) Immunofluorescence images of DAPI (blue)-CD31 (red) HBMECs co-cultured with CM-Dio (green) labeled ncU87-Exo and shU87-Exo at 6 h. (B) qRT-PCR was applied to determine linc-CCAT2 expression levels in HUVECs when incubated with 100 µg/ml ncU87-Exo and shU87-Exo for 24 h; the linc-CCAT2 expression level in ncU87-Exo-treated HUVECs was significantly higher than that in the shU87-Exo-treated HUVECs (*P

    Journal: Oncology Reports

    Article Title: Glioma cells enhance angiogenesis and inhibit endothelial cell apoptosis through the release of exosomes that contain long non-coding RNA CCAT2

    doi: 10.3892/or.2017.5742

    Figure Lengend Snippet: ncU87-Exo and shU87-Exo are internalized by HUVECs. (A) Immunofluorescence images of DAPI (blue)-CD31 (red) HBMECs co-cultured with CM-Dio (green) labeled ncU87-Exo and shU87-Exo at 6 h. (B) qRT-PCR was applied to determine linc-CCAT2 expression levels in HUVECs when incubated with 100 µg/ml ncU87-Exo and shU87-Exo for 24 h; the linc-CCAT2 expression level in ncU87-Exo-treated HUVECs was significantly higher than that in the shU87-Exo-treated HUVECs (*P

    Article Snippet: RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR) assay RNAzol RT (Molecular Research Center, USA) was used to extract total RNA from glioma cell lines, HUVECs and exosomal sources ( ). qRT-PCR for human-specific repeat sequences was performed as previously described ( ).

    Techniques: Immunofluorescence, Cell Culture, Labeling, Quantitative RT-PCR, Expressing, Incubation

    ncU87-Exo and shU87-Exo regulate the expression of apoptosis-related factors in HUVECs induced by hypoxia. (A) qRT-PCR analysis of the expression level of apoptosis-related factors Bcl-2, Bax, and caspase-3 in HUVECs treated by ncU87-Exo and shU87-Exo after hypoxia. Compared with the control medium group and the shU87-Exo group, ncU87-Exo significantly upregulated Bcl-2 gene expression and inhibited Bax and caspase-3 gene expression. (B and C) Western blot analysis revealed that both ncU87-Exo and shU87-Exo increased Bcl-2 expression and decreased Bax and caspase-3 expression, while ncU87-Exo was more efficient. (*P

    Journal: Oncology Reports

    Article Title: Glioma cells enhance angiogenesis and inhibit endothelial cell apoptosis through the release of exosomes that contain long non-coding RNA CCAT2

    doi: 10.3892/or.2017.5742

    Figure Lengend Snippet: ncU87-Exo and shU87-Exo regulate the expression of apoptosis-related factors in HUVECs induced by hypoxia. (A) qRT-PCR analysis of the expression level of apoptosis-related factors Bcl-2, Bax, and caspase-3 in HUVECs treated by ncU87-Exo and shU87-Exo after hypoxia. Compared with the control medium group and the shU87-Exo group, ncU87-Exo significantly upregulated Bcl-2 gene expression and inhibited Bax and caspase-3 gene expression. (B and C) Western blot analysis revealed that both ncU87-Exo and shU87-Exo increased Bcl-2 expression and decreased Bax and caspase-3 expression, while ncU87-Exo was more efficient. (*P

    Article Snippet: RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR) assay RNAzol RT (Molecular Research Center, USA) was used to extract total RNA from glioma cell lines, HUVECs and exosomal sources ( ). qRT-PCR for human-specific repeat sequences was performed as previously described ( ).

    Techniques: Expressing, Quantitative RT-PCR, Western Blot

    ncU87-Exo and shU87-Exo regulate angiogenesis related-genes and protein expression in HUVECs. (A) qRT-PCR analysis of the expression level of angiogenesis-related genes in HUVECs treated by ncU87-Exo and shU87-Exo. Compared with the shU87-Exo group, ncU87-Exo significantly upregulated VEGF, TGFβ, FGF and KDR gene expression. (B) ELISA analysis of the secretion level of angiogenesis-related proteins in HUVECs treated by ncU87-Exo and shU87-Exo. Compared with the shU87-Exo group, ncU87-Exo significantly increased VEGF, TGFβ, and FGF protein secretion. (*P

    Journal: Oncology Reports

    Article Title: Glioma cells enhance angiogenesis and inhibit endothelial cell apoptosis through the release of exosomes that contain long non-coding RNA CCAT2

    doi: 10.3892/or.2017.5742

    Figure Lengend Snippet: ncU87-Exo and shU87-Exo regulate angiogenesis related-genes and protein expression in HUVECs. (A) qRT-PCR analysis of the expression level of angiogenesis-related genes in HUVECs treated by ncU87-Exo and shU87-Exo. Compared with the shU87-Exo group, ncU87-Exo significantly upregulated VEGF, TGFβ, FGF and KDR gene expression. (B) ELISA analysis of the secretion level of angiogenesis-related proteins in HUVECs treated by ncU87-Exo and shU87-Exo. Compared with the shU87-Exo group, ncU87-Exo significantly increased VEGF, TGFβ, and FGF protein secretion. (*P

    Article Snippet: RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR) assay RNAzol RT (Molecular Research Center, USA) was used to extract total RNA from glioma cell lines, HUVECs and exosomal sources ( ). qRT-PCR for human-specific repeat sequences was performed as previously described ( ).

    Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    linc-CCAT2 regulates HUVEC migration, proliferation, and tube formation in vitro . (A) The linc-CCAT2 expression levels were evaluated by qRT-PCR in linc-CCAT2 overexpression, downregulation, negative control, and normal HUVECs. linc-CCAT2 overexpression in HUVECs exhibited the highest expression level of linc-CCAT2, while downregulation of linc-CCAT2 in HUVECs exhibited the lowest expression level of linc-CCAT2. (B and C) The migration ability was assessed with the scratch-wound assay; linc-CCAT2 overexpression increased HUVEC migration, while downregulation of linc-CCAT2 significantly decreased HUVEC migration. (D) The proliferation was assessed with the CCK-8 assay; linc-CCAT2 overexpression promoted HUVEC proliferation, while linc-CCAT2 downexpression inhibited HUVEC proliferation. (E-H) The tube formation assay was performed on growth factor-reduced Matrigel. The total number of branching points, total tube length, and total loops at 6 h in the linc-CCAT2 overexpression in HUVECs was higher than in the negative control HUVECs, while downregulation of linc-CCAT2 in HUVECs was lower than the negative control HUVECs. (*P

    Journal: Oncology Reports

    Article Title: Glioma cells enhance angiogenesis and inhibit endothelial cell apoptosis through the release of exosomes that contain long non-coding RNA CCAT2

    doi: 10.3892/or.2017.5742

    Figure Lengend Snippet: linc-CCAT2 regulates HUVEC migration, proliferation, and tube formation in vitro . (A) The linc-CCAT2 expression levels were evaluated by qRT-PCR in linc-CCAT2 overexpression, downregulation, negative control, and normal HUVECs. linc-CCAT2 overexpression in HUVECs exhibited the highest expression level of linc-CCAT2, while downregulation of linc-CCAT2 in HUVECs exhibited the lowest expression level of linc-CCAT2. (B and C) The migration ability was assessed with the scratch-wound assay; linc-CCAT2 overexpression increased HUVEC migration, while downregulation of linc-CCAT2 significantly decreased HUVEC migration. (D) The proliferation was assessed with the CCK-8 assay; linc-CCAT2 overexpression promoted HUVEC proliferation, while linc-CCAT2 downexpression inhibited HUVEC proliferation. (E-H) The tube formation assay was performed on growth factor-reduced Matrigel. The total number of branching points, total tube length, and total loops at 6 h in the linc-CCAT2 overexpression in HUVECs was higher than in the negative control HUVECs, while downregulation of linc-CCAT2 in HUVECs was lower than the negative control HUVECs. (*P

    Article Snippet: RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR) assay RNAzol RT (Molecular Research Center, USA) was used to extract total RNA from glioma cell lines, HUVECs and exosomal sources ( ). qRT-PCR for human-specific repeat sequences was performed as previously described ( ).

    Techniques: Migration, In Vitro, Expressing, Quantitative RT-PCR, Over Expression, Negative Control, Scratch Wound Assay Assay, CCK-8 Assay, Tube Formation Assay

    linc-CCAT2 regulates apoptosis-related factor expression in HUVECs induced by hypoxia. (A) qRT-PCR analysis of the expression level of apoptosis-related factors Bcl-2, Bax and caspase-3 in linc-CCAT2 overexpression, downregulation and negative control HUVECs. Compared with the negative control HUVECs, overexpression promoted Bcl-2 gene expression and inhibited Bax and caspase-3 gene expression, while downregulation of linc-CCAT2 in HUVECs inhibited Bcl-2 gene expression and promoted Bax and caspase-3 gene expression. (B and C) Western blot analysis of the expression level of Bcl-2, Bax, and caspase-3 in HUVECs. Compared with the negative control HUVECs, linc-CCAT2 overexpression improved Bcl-2 protein expression and decreased Bax and caspase-3 protein expression, while downregulation of linc-CCAT2 decreased Bcl-2 protein expression and increased Bax and caspase-3 protein expression. (*P

    Journal: Oncology Reports

    Article Title: Glioma cells enhance angiogenesis and inhibit endothelial cell apoptosis through the release of exosomes that contain long non-coding RNA CCAT2

    doi: 10.3892/or.2017.5742

    Figure Lengend Snippet: linc-CCAT2 regulates apoptosis-related factor expression in HUVECs induced by hypoxia. (A) qRT-PCR analysis of the expression level of apoptosis-related factors Bcl-2, Bax and caspase-3 in linc-CCAT2 overexpression, downregulation and negative control HUVECs. Compared with the negative control HUVECs, overexpression promoted Bcl-2 gene expression and inhibited Bax and caspase-3 gene expression, while downregulation of linc-CCAT2 in HUVECs inhibited Bcl-2 gene expression and promoted Bax and caspase-3 gene expression. (B and C) Western blot analysis of the expression level of Bcl-2, Bax, and caspase-3 in HUVECs. Compared with the negative control HUVECs, linc-CCAT2 overexpression improved Bcl-2 protein expression and decreased Bax and caspase-3 protein expression, while downregulation of linc-CCAT2 decreased Bcl-2 protein expression and increased Bax and caspase-3 protein expression. (*P

    Article Snippet: RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR) assay RNAzol RT (Molecular Research Center, USA) was used to extract total RNA from glioma cell lines, HUVECs and exosomal sources ( ). qRT-PCR for human-specific repeat sequences was performed as previously described ( ).

    Techniques: Expressing, Quantitative RT-PCR, Over Expression, Negative Control, Western Blot

    Characterization of exosomes released by ncU87-MG cells and shU87-MG cells. (A) linc-CCAT2 expression levels were evaluated by qRT-PCR in the following four glioma cell lines: U87-MG, U251, A172, and T98G; non-glioma 293T cells were used as controls. U87-MG cells exhibited the highest expression level of linc-CCAT2 (compared with the 293T cells, *P

    Journal: Oncology Reports

    Article Title: Glioma cells enhance angiogenesis and inhibit endothelial cell apoptosis through the release of exosomes that contain long non-coding RNA CCAT2

    doi: 10.3892/or.2017.5742

    Figure Lengend Snippet: Characterization of exosomes released by ncU87-MG cells and shU87-MG cells. (A) linc-CCAT2 expression levels were evaluated by qRT-PCR in the following four glioma cell lines: U87-MG, U251, A172, and T98G; non-glioma 293T cells were used as controls. U87-MG cells exhibited the highest expression level of linc-CCAT2 (compared with the 293T cells, *P

    Article Snippet: RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR) assay RNAzol RT (Molecular Research Center, USA) was used to extract total RNA from glioma cell lines, HUVECs and exosomal sources ( ). qRT-PCR for human-specific repeat sequences was performed as previously described ( ).

    Techniques: Expressing, Quantitative RT-PCR

    linc-CCAT2 regulates angiogenesis-related genes and protein expression in HUVECs. (A) qRT-PCR analysis of the expression level of angiogenesis-related genes in linc-CCAT2 overexpression, downregulation and negative control HUVECs. linc-CCAT2 overexpression in HUVECs increased the gene expression of VEGF, TGFβ, and KDR, while downregulation of linc-CCAT2 inhibited VEGF, TGFβ, and KDR expression. (B) ELISA analysis of the level of secreted angiogenesis-related proteins in linc-CCAT2 overexpression, downregulation and negative control HUVECs. linc-CCAT2 overexpression in HUVECs increased VEGF and TGFβ protein secretion, while downregulation of linc-CCAT2 inhibited VEGF and TGFβ protein secretion. (*P

    Journal: Oncology Reports

    Article Title: Glioma cells enhance angiogenesis and inhibit endothelial cell apoptosis through the release of exosomes that contain long non-coding RNA CCAT2

    doi: 10.3892/or.2017.5742

    Figure Lengend Snippet: linc-CCAT2 regulates angiogenesis-related genes and protein expression in HUVECs. (A) qRT-PCR analysis of the expression level of angiogenesis-related genes in linc-CCAT2 overexpression, downregulation and negative control HUVECs. linc-CCAT2 overexpression in HUVECs increased the gene expression of VEGF, TGFβ, and KDR, while downregulation of linc-CCAT2 inhibited VEGF, TGFβ, and KDR expression. (B) ELISA analysis of the level of secreted angiogenesis-related proteins in linc-CCAT2 overexpression, downregulation and negative control HUVECs. linc-CCAT2 overexpression in HUVECs increased VEGF and TGFβ protein secretion, while downregulation of linc-CCAT2 inhibited VEGF and TGFβ protein secretion. (*P

    Article Snippet: RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR) assay RNAzol RT (Molecular Research Center, USA) was used to extract total RNA from glioma cell lines, HUVECs and exosomal sources ( ). qRT-PCR for human-specific repeat sequences was performed as previously described ( ).

    Techniques: Expressing, Quantitative RT-PCR, Over Expression, Negative Control, Enzyme-linked Immunosorbent Assay

    Comparison of difference of miRNA-19a expression between model and normal group. The contents of miRNA-19a in rat gastric tissue of model and normal group are detected by qRT-PCR, which displayed that the relative content of miRNA-19a in gastric tissue of model group is 6.13±0.89, which is 4.56±0.65 in normal tissue, suggesting that miRNA-19a is highly expressed in rats with functional dyspepsia, *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Effect of miRNA-19a on gastrointestinal motility in rats with functional dyspepsia

    doi: 10.3892/etm.2018.6009

    Figure Lengend Snippet: Comparison of difference of miRNA-19a expression between model and normal group. The contents of miRNA-19a in rat gastric tissue of model and normal group are detected by qRT-PCR, which displayed that the relative content of miRNA-19a in gastric tissue of model group is 6.13±0.89, which is 4.56±0.65 in normal tissue, suggesting that miRNA-19a is highly expressed in rats with functional dyspepsia, *P

    Article Snippet: TRIzol, reverse-transcription kit and real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) kit (Promega Corp., Madison, WI, USA), miRNA-19a, U6 specific primers (Shanghai Sangon Biological Engineering Co., Ltd., Shanghai, China), miRNA-19a sequence mimics (miRNA-19a mimic) and miRNA-19a unrelated sequence mimics (scramble mimic) (Shanghai Sangon Biological Engineering Co., Ltd.), In vivo -jetPEI® (Polyplus Transfection, Illkirch, France), rat motilin (MTL), vasoactive intestinal peptide (VIP) (Wuhan Huamei Biological Engineering Co., Ltd., Wuhan, China), real-time fluorescent quantitative PCR 7500 system (Applied Biosystems, Foster City, CA, USA).

    Techniques: Expressing, Quantitative RT-PCR, Functional Assay