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Bio-Rad total ribonucleic acid rna
Total Ribonucleic Acid Rna, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/total ribonucleic acid rna/product/Bio-Rad
Average 92 stars, based on 2 article reviews
Price from $9.99 to $1999.99
total ribonucleic acid rna - by Bioz Stars, 2020-04
92/100 stars

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Related Articles

Real-time Polymerase Chain Reaction:

Article Title: Chemokine (C‐C Motif) Receptor‐Like 2 is not essential for lung injury, lung inflammation, or airway hyperresponsiveness induced by acute exposure to ozone. Chemokine (C‐C Motif) Receptor‐like 2 is not essential for lung injury, lung inflammation, or airway hyperresponsiveness induced by acute exposure to ozone
Article Snippet: .. At a later date, total ribonucleic acid (RNA) was extracted from the left lung lobe and complementary deoxyribonucleic acid was synthesized from messenger RNA (mRNA) as previously described (Razvi et al. ). qPCR was performed to determine the relative abundance of Ccrl2 mRNA in the left lung lobe using iTaq™ Universal SYBR® Green Supermix (Bio‐Rad Laboratories, Inc.; Hercules, CA) and a CFX Connect™ Real‐Time PCR Detection System (Bio‐Rad Laboratories, Inc.) as per the instructions of the manufacturer. ..

Article Title: Osteogenic potential of sorted equine mesenchymal stem cell subpopulations
Article Snippet: Paragraph title: Real-time quantitative PCR ... Growth of the paired cultures was stopped on day 7 and total ribonucleic acid (RNA) was extracted (Aurum Total RNA Mini Kit, Bio-Rad Laboratories, Hercules, California, USA) from the cells.

Synthesized:

Article Title: Chemokine (C‐C Motif) Receptor‐Like 2 is not essential for lung injury, lung inflammation, or airway hyperresponsiveness induced by acute exposure to ozone. Chemokine (C‐C Motif) Receptor‐like 2 is not essential for lung injury, lung inflammation, or airway hyperresponsiveness induced by acute exposure to ozone
Article Snippet: .. At a later date, total ribonucleic acid (RNA) was extracted from the left lung lobe and complementary deoxyribonucleic acid was synthesized from messenger RNA (mRNA) as previously described (Razvi et al. ). qPCR was performed to determine the relative abundance of Ccrl2 mRNA in the left lung lobe using iTaq™ Universal SYBR® Green Supermix (Bio‐Rad Laboratories, Inc.; Hercules, CA) and a CFX Connect™ Real‐Time PCR Detection System (Bio‐Rad Laboratories, Inc.) as per the instructions of the manufacturer. ..

Article Title: Osteogenic potential of sorted equine mesenchymal stem cell subpopulations
Article Snippet: Growth of the paired cultures was stopped on day 7 and total ribonucleic acid (RNA) was extracted (Aurum Total RNA Mini Kit, Bio-Rad Laboratories, Hercules, California, USA) from the cells. .. The complementary deoxyribonucleic acid (cDNA) was synthesized from total RNA via a cDNA synthesis kit (iScript cDNA Synthesis Kit; Bio-Rad Laboratories).

Cell Culture:

Article Title: Osteogenic potential of sorted equine mesenchymal stem cell subpopulations
Article Snippet: Cultured and expanded cells from the second passage of each of the 5 fractions and the 1 unsorted control from both bone marrow and muscle tissue were seeded in 6-well plates at 200 cells/cm2 in triplicate. .. Growth of the paired cultures was stopped on day 7 and total ribonucleic acid (RNA) was extracted (Aurum Total RNA Mini Kit, Bio-Rad Laboratories, Hercules, California, USA) from the cells.

Mouse Assay:

Article Title: Chemokine (C‐C Motif) Receptor‐Like 2 is not essential for lung injury, lung inflammation, or airway hyperresponsiveness induced by acute exposure to ozone. Chemokine (C‐C Motif) Receptor‐like 2 is not essential for lung injury, lung inflammation, or airway hyperresponsiveness induced by acute exposure to ozone
Article Snippet: Reverse transcription (RT)‐quantitative real‐time polymerase chain reactions (qPCR) Following the collection of blood from wild‐type mice that were part of the first cohort, the left lung lobe of these animals was removed by severing the left main bronchus. .. At a later date, total ribonucleic acid (RNA) was extracted from the left lung lobe and complementary deoxyribonucleic acid was synthesized from messenger RNA (mRNA) as previously described (Razvi et al. ). qPCR was performed to determine the relative abundance of Ccrl2 mRNA in the left lung lobe using iTaq™ Universal SYBR® Green Supermix (Bio‐Rad Laboratories, Inc.; Hercules, CA) and a CFX Connect™ Real‐Time PCR Detection System (Bio‐Rad Laboratories, Inc.) as per the instructions of the manufacturer.

SYBR Green Assay:

Article Title: Chemokine (C‐C Motif) Receptor‐Like 2 is not essential for lung injury, lung inflammation, or airway hyperresponsiveness induced by acute exposure to ozone. Chemokine (C‐C Motif) Receptor‐like 2 is not essential for lung injury, lung inflammation, or airway hyperresponsiveness induced by acute exposure to ozone
Article Snippet: .. At a later date, total ribonucleic acid (RNA) was extracted from the left lung lobe and complementary deoxyribonucleic acid was synthesized from messenger RNA (mRNA) as previously described (Razvi et al. ). qPCR was performed to determine the relative abundance of Ccrl2 mRNA in the left lung lobe using iTaq™ Universal SYBR® Green Supermix (Bio‐Rad Laboratories, Inc.; Hercules, CA) and a CFX Connect™ Real‐Time PCR Detection System (Bio‐Rad Laboratories, Inc.) as per the instructions of the manufacturer. ..

Derivative Assay:

Article Title: Osteogenic potential of sorted equine mesenchymal stem cell subpopulations
Article Snippet: Growth of the paired cultures was stopped on day 7 and total ribonucleic acid (RNA) was extracted (Aurum Total RNA Mini Kit, Bio-Rad Laboratories, Hercules, California, USA) from the cells. .. Primers derived from the coding regions of osteocalcin (OCN) were as follows: forward, 5′-CTGGGCCAGGACTCCGCATCT-3′ and reverse, 5′-AGCCAGCTCGTCACAGTCTGGGTTG-3′.

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    98
    Bio-Rad rna
    Pregnancy promotes hepatic glucocorticoid resistance via receptor downregulation. a Whole-transcriptome array of livers from virgins and mice pregnant at 14.5 dpc overlaid with previously published <t>RNA-seq</t> of glucocorticoid-responsive genes in the virgin female liver (GSE99309). b Top physiological pathways activated in the pregnant liver compared with glucocorticoid regulation of those pathways. c mRNA expression of GR target genes in pregnant liver. n = 4–7 mice per group; df = 33. d Real-time <t>PCR</t> analysis of livers from dexamethasone challenge (10 μg/kg) in female virgins and pregnant mice. n = 3 mice per group; df = 11. e Immunoblot and quantification of GR protein in livers from virgin and pregnant mice. β-actin was used as a loading control and for normalization. n = 9–10 animals per group; df = 17. f NR3C1 mRNA in virgin and pregnant livers. n = 4–7 mice per group; df = 9. g Expression of nascent NR3C1 mRNAs from virgin and pregnant livers. n = 4 mice per group; df = 6. Data are expressed as mean ± SEM. * denotes p
    Rna, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 98/100, based on 721 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rna/product/Bio-Rad
    Average 98 stars, based on 721 article reviews
    Price from $9.99 to $1999.99
    rna - by Bioz Stars, 2020-04
    98/100 stars
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    94
    Bio-Rad quantitative real time reverse transcription rt pcr purified total rna
    Pearson correlation value between the log2-transformed fold changes of <t>RNA-Seq</t> data and <t>qRT-PCR</t> validation. Each dot (in blue) represents a data point for the selected genes and each corresponding sample (4 genes and 8 replicates in control (D0) and later time point, D1 and D14, respectively). And the dashed lines are 95% confidence interval.
    Quantitative Real Time Reverse Transcription Rt Pcr Purified Total Rna, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time reverse transcription rt pcr purified total rna/product/Bio-Rad
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    quantitative real time reverse transcription rt pcr purified total rna - by Bioz Stars, 2020-04
    94/100 stars
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    93
    Bio-Rad real time pcr analysis total rna
    High temperature induces endoplasmic reticulum (ER) stress-mediated cell death. ( a ) Protein expression under condition of hyperthermia was measured. Cell lysates were collected at indicated times following incubation at 42 °C or tunicamycin (Tm) (1 μg/mL) treatment for western blot analysis. ( b ) Effect of heat stress on the unfolded protein response (UPR) gene expression. Quantitative <t>RT-PCR</t> was performed using total <t>RNA</t> extracted from mouse embryonic fibroblasts (MEFs) which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( c ) KDEL and DAPI immunofluorescence staining. KDEL and DAPI staining were performed using MEFs incubated at 42 °C or treated with thapsigargin (Tg) (300 nM) for 12 h. Representative images are shown. Scale bar represents 100 μm. ( d ) Cell viability was measured in MEFs treated with Tg (300 nM), Tm (1 μg/mL), or incubated at 42 °C for the indicated time periods. ( e ) Cell viability was measured in MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of tauroursodeoxycholic acid (TUDCA) (2.4 mM). ** p
    Real Time Pcr Analysis Total Rna, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time pcr analysis total rna/product/Bio-Rad
    Average 93 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    real time pcr analysis total rna - by Bioz Stars, 2020-04
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    Pregnancy promotes hepatic glucocorticoid resistance via receptor downregulation. a Whole-transcriptome array of livers from virgins and mice pregnant at 14.5 dpc overlaid with previously published RNA-seq of glucocorticoid-responsive genes in the virgin female liver (GSE99309). b Top physiological pathways activated in the pregnant liver compared with glucocorticoid regulation of those pathways. c mRNA expression of GR target genes in pregnant liver. n = 4–7 mice per group; df = 33. d Real-time PCR analysis of livers from dexamethasone challenge (10 μg/kg) in female virgins and pregnant mice. n = 3 mice per group; df = 11. e Immunoblot and quantification of GR protein in livers from virgin and pregnant mice. β-actin was used as a loading control and for normalization. n = 9–10 animals per group; df = 17. f NR3C1 mRNA in virgin and pregnant livers. n = 4–7 mice per group; df = 9. g Expression of nascent NR3C1 mRNAs from virgin and pregnant livers. n = 4 mice per group; df = 6. Data are expressed as mean ± SEM. * denotes p

    Journal: Communications Biology

    Article Title: Silencing of maternal hepatic glucocorticoid receptor is essential for normal fetal development in mice

    doi: 10.1038/s42003-019-0344-3

    Figure Lengend Snippet: Pregnancy promotes hepatic glucocorticoid resistance via receptor downregulation. a Whole-transcriptome array of livers from virgins and mice pregnant at 14.5 dpc overlaid with previously published RNA-seq of glucocorticoid-responsive genes in the virgin female liver (GSE99309). b Top physiological pathways activated in the pregnant liver compared with glucocorticoid regulation of those pathways. c mRNA expression of GR target genes in pregnant liver. n = 4–7 mice per group; df = 33. d Real-time PCR analysis of livers from dexamethasone challenge (10 μg/kg) in female virgins and pregnant mice. n = 3 mice per group; df = 11. e Immunoblot and quantification of GR protein in livers from virgin and pregnant mice. β-actin was used as a loading control and for normalization. n = 9–10 animals per group; df = 17. f NR3C1 mRNA in virgin and pregnant livers. n = 4–7 mice per group; df = 9. g Expression of nascent NR3C1 mRNAs from virgin and pregnant livers. n = 4 mice per group; df = 6. Data are expressed as mean ± SEM. * denotes p

    Article Snippet: Quantitative PCR One-hundred nanograms of the total RNA were reverse-transcribed and amplified according to the manufacturer’s instructions for the iScript One-Step RT-PCR kit for probes (Biorad; Hercules, CA).

    Techniques: Mouse Assay, RNA Sequencing Assay, Expressing, Real-time Polymerase Chain Reaction

    In vitro and in vivo gene expression analysis of selected Mucor genes. (A) RT-qPCR-determined differences in the expression of selected Mucor genes during mouse macrophage phagocytosis. Transcript levels were quantified in cDNA obtained from virulent Mucor strain R7B spores cocultured with mouse macrophages (Mφ) (cell line J774A.1) in L15 cell culture medium for 5 h and from the same spores cultured in noninteraction controls (without macrophages). Values were normalized using 18S rRNA as an internal control, and the differential expression (log 2 -fold change) was calculated with respect to expression in noninteraction samples. (B) RT-qPCR-determined differences in the expression of the selected genes during interaction with peritoneal macrophages. Transcript levels were quantified in cDNA obtained from virulent Mucor strain R7B spores that were injected into the peritoneal cavity of OF-1 mice and recovered after 5 h, as well as from the same spores incubated in L15 cell culture medium as a control. Values were normalized using 18S rRNA as an internal control, and the differential expression (log 2 -fold change) was calculated with respect to the expression in L15 control samples. (C) Comparison of differential expression of selected genes during in vitro interaction with mouse macrophages, quantified by both RNA-seq and RT-qPCR, and during in vivo interaction with peritoneal macrophages, quantified by RT-qPCR. Error bars correspond to the standard deviations (SD) of the results from technical triplicates, and asterisks indicate a significant difference determined by unpaired t test (*, P ≤ 0.05; **, P ≤ 0.005; ***, P

    Journal: mBio

    Article Title: Mucor circinelloides Thrives inside the Phagosome through an Atf-Mediated Germination Pathway

    doi: 10.1128/mBio.02765-18

    Figure Lengend Snippet: In vitro and in vivo gene expression analysis of selected Mucor genes. (A) RT-qPCR-determined differences in the expression of selected Mucor genes during mouse macrophage phagocytosis. Transcript levels were quantified in cDNA obtained from virulent Mucor strain R7B spores cocultured with mouse macrophages (Mφ) (cell line J774A.1) in L15 cell culture medium for 5 h and from the same spores cultured in noninteraction controls (without macrophages). Values were normalized using 18S rRNA as an internal control, and the differential expression (log 2 -fold change) was calculated with respect to expression in noninteraction samples. (B) RT-qPCR-determined differences in the expression of the selected genes during interaction with peritoneal macrophages. Transcript levels were quantified in cDNA obtained from virulent Mucor strain R7B spores that were injected into the peritoneal cavity of OF-1 mice and recovered after 5 h, as well as from the same spores incubated in L15 cell culture medium as a control. Values were normalized using 18S rRNA as an internal control, and the differential expression (log 2 -fold change) was calculated with respect to the expression in L15 control samples. (C) Comparison of differential expression of selected genes during in vitro interaction with mouse macrophages, quantified by both RNA-seq and RT-qPCR, and during in vivo interaction with peritoneal macrophages, quantified by RT-qPCR. Error bars correspond to the standard deviations (SD) of the results from technical triplicates, and asterisks indicate a significant difference determined by unpaired t test (*, P ≤ 0.05; **, P ≤ 0.005; ***, P

    Article Snippet: For RT-qPCR assays, cDNA was synthesized from 1 µg of total RNA using the iScript cDNA synthesis kit (Bio-Rad).

    Techniques: In Vitro, In Vivo, Expressing, Quantitative RT-PCR, Cell Culture, Injection, Mouse Assay, Incubation, RNA Sequencing Assay

    Pearson correlation value between the log2-transformed fold changes of RNA-Seq data and qRT-PCR validation. Each dot (in blue) represents a data point for the selected genes and each corresponding sample (4 genes and 8 replicates in control (D0) and later time point, D1 and D14, respectively). And the dashed lines are 95% confidence interval.

    Journal: Scientific Reports

    Article Title: Temporal dynamics in meta longitudinal RNA-Seq data

    doi: 10.1038/s41598-018-37397-7

    Figure Lengend Snippet: Pearson correlation value between the log2-transformed fold changes of RNA-Seq data and qRT-PCR validation. Each dot (in blue) represents a data point for the selected genes and each corresponding sample (4 genes and 8 replicates in control (D0) and later time point, D1 and D14, respectively). And the dashed lines are 95% confidence interval.

    Article Snippet: Quantitative Real-time Reverse Transcription (RT) PCR Purified total RNA samples were converted to cDNA using an iScript Advanced cDNA Synthesis Kit (Bio-Rad, Hercules, CA, USA).

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

    High temperature induces endoplasmic reticulum (ER) stress-mediated cell death. ( a ) Protein expression under condition of hyperthermia was measured. Cell lysates were collected at indicated times following incubation at 42 °C or tunicamycin (Tm) (1 μg/mL) treatment for western blot analysis. ( b ) Effect of heat stress on the unfolded protein response (UPR) gene expression. Quantitative RT-PCR was performed using total RNA extracted from mouse embryonic fibroblasts (MEFs) which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( c ) KDEL and DAPI immunofluorescence staining. KDEL and DAPI staining were performed using MEFs incubated at 42 °C or treated with thapsigargin (Tg) (300 nM) for 12 h. Representative images are shown. Scale bar represents 100 μm. ( d ) Cell viability was measured in MEFs treated with Tg (300 nM), Tm (1 μg/mL), or incubated at 42 °C for the indicated time periods. ( e ) Cell viability was measured in MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of tauroursodeoxycholic acid (TUDCA) (2.4 mM). ** p

    Journal: Cells

    Article Title: Modulation of Protein Synthesis by eIF2α Phosphorylation Protects Cell from Heat Stress-Mediated Apoptosis

    doi: 10.3390/cells7120254

    Figure Lengend Snippet: High temperature induces endoplasmic reticulum (ER) stress-mediated cell death. ( a ) Protein expression under condition of hyperthermia was measured. Cell lysates were collected at indicated times following incubation at 42 °C or tunicamycin (Tm) (1 μg/mL) treatment for western blot analysis. ( b ) Effect of heat stress on the unfolded protein response (UPR) gene expression. Quantitative RT-PCR was performed using total RNA extracted from mouse embryonic fibroblasts (MEFs) which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( c ) KDEL and DAPI immunofluorescence staining. KDEL and DAPI staining were performed using MEFs incubated at 42 °C or treated with thapsigargin (Tg) (300 nM) for 12 h. Representative images are shown. Scale bar represents 100 μm. ( d ) Cell viability was measured in MEFs treated with Tg (300 nM), Tm (1 μg/mL), or incubated at 42 °C for the indicated time periods. ( e ) Cell viability was measured in MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of tauroursodeoxycholic acid (TUDCA) (2.4 mM). ** p

    Article Snippet: RNA Extraction and Real-Time PCR Analysis Total RNA was extracted from cells using Trizol (Gene All, Ribo-Ex, Seoul, Korea) and cDNA was synthesized using an iScript cDNA synthesis kit (Bio-Rad, Hercules, CA, USA, BR170-8891).

    Techniques: Expressing, Incubation, Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining

    The ATF6α pathway does not protect cells from heat stress-mediated death. ( a ) Quantitative RT-PCR was performed using total RNA extracted from Atf6α −/− and Atf6α +/+ MEFs which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( b ) Protein expression under conditions of hyperthermia was measured. Following 42 °C incubation, cell lysates were collected at indicated times for western blot analysis. ( c ) Cell viability was measured in Atf6α −/− and Atf6α +/+ MEFs following incubation at 42 °C for indicated time periods. ( d ) Caspase 3/7 activity was detected in Atf6α −/− and Atf6α +/+ MEFs which were incubated at 42 °C for 12 h. ** p

    Journal: Cells

    Article Title: Modulation of Protein Synthesis by eIF2α Phosphorylation Protects Cell from Heat Stress-Mediated Apoptosis

    doi: 10.3390/cells7120254

    Figure Lengend Snippet: The ATF6α pathway does not protect cells from heat stress-mediated death. ( a ) Quantitative RT-PCR was performed using total RNA extracted from Atf6α −/− and Atf6α +/+ MEFs which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( b ) Protein expression under conditions of hyperthermia was measured. Following 42 °C incubation, cell lysates were collected at indicated times for western blot analysis. ( c ) Cell viability was measured in Atf6α −/− and Atf6α +/+ MEFs following incubation at 42 °C for indicated time periods. ( d ) Caspase 3/7 activity was detected in Atf6α −/− and Atf6α +/+ MEFs which were incubated at 42 °C for 12 h. ** p

    Article Snippet: RNA Extraction and Real-Time PCR Analysis Total RNA was extracted from cells using Trizol (Gene All, Ribo-Ex, Seoul, Korea) and cDNA was synthesized using an iScript cDNA synthesis kit (Bio-Rad, Hercules, CA, USA, BR170-8891).

    Techniques: Quantitative RT-PCR, Incubation, Expressing, Western Blot, Activity Assay

    eIF2α phosphorylation is required to protect cells from heat stress-mediated death. ( a ) Protein expression was measured under conditions of hyperthermia. Cell lysates were collected from Eif2α S/S and Eif2α A/A MEFs at indicated times for western blot analysis following incubation at 42 °C. ( b ) Quantitative RT-PCR was performed using total RNA extracted from Eif2α S/S and Eif2α A/A MEFs which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( c ) Cell viability was measured in Eif2α S/S and Eif2α A/A MEFs which were incubated at 42 °C for indicated times. ( d ) Caspase 3/7 activity was detected in Eif2α S/S and Eif2α A/A MEFs which were incubated at 42 °C for 12 h. ( e ) Cell viability was measured in MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of guanabenz (GA, 1 μM) or ISRIB (5 μM). ( f ) Cell viability was measured in Perk +/+ and Perk −/− MEFs which were incubated at 42 °C for 12 and 24 h. ( g ) Cell viability was measured in MEFs which were incubated at 42 °C for 12 h in the presence or absence of GSK2606414 (100 nM). * p

    Journal: Cells

    Article Title: Modulation of Protein Synthesis by eIF2α Phosphorylation Protects Cell from Heat Stress-Mediated Apoptosis

    doi: 10.3390/cells7120254

    Figure Lengend Snippet: eIF2α phosphorylation is required to protect cells from heat stress-mediated death. ( a ) Protein expression was measured under conditions of hyperthermia. Cell lysates were collected from Eif2α S/S and Eif2α A/A MEFs at indicated times for western blot analysis following incubation at 42 °C. ( b ) Quantitative RT-PCR was performed using total RNA extracted from Eif2α S/S and Eif2α A/A MEFs which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( c ) Cell viability was measured in Eif2α S/S and Eif2α A/A MEFs which were incubated at 42 °C for indicated times. ( d ) Caspase 3/7 activity was detected in Eif2α S/S and Eif2α A/A MEFs which were incubated at 42 °C for 12 h. ( e ) Cell viability was measured in MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of guanabenz (GA, 1 μM) or ISRIB (5 μM). ( f ) Cell viability was measured in Perk +/+ and Perk −/− MEFs which were incubated at 42 °C for 12 and 24 h. ( g ) Cell viability was measured in MEFs which were incubated at 42 °C for 12 h in the presence or absence of GSK2606414 (100 nM). * p

    Article Snippet: RNA Extraction and Real-Time PCR Analysis Total RNA was extracted from cells using Trizol (Gene All, Ribo-Ex, Seoul, Korea) and cDNA was synthesized using an iScript cDNA synthesis kit (Bio-Rad, Hercules, CA, USA, BR170-8891).

    Techniques: Expressing, Western Blot, Incubation, Quantitative RT-PCR, Activity Assay

    The IRE1α pathway does not protect cells from heat stress-mediated death. ( a ) Quantitative RT-PCR was performed using total RNA, extracted from Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM. (n = 3 independent experiments). ( b ) Protein expression under conditions of hyperthermia was measured. Following incubation at 42 °C, cell lysates were collected at the indicated times for western blot analysis. ( c ) Cell viability was measured in Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for indicated time periods. ( d ) Caspase 3/7 activity was detected in Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for 12 h. ( e ) Expression of sXBP1 (spliced form) and tXBP (total form) were measured using MEFs treated with 4μ8c at indicated doses in the presence or absence of Tg (300 nM) for 12 h. ( f ) Protein expression under conditions of hyperthermia was measured in the presence or absence of 4μ8c (20 μM). ( g ) Cell viability was measured in Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of 4μ8c (20 μM). * p

    Journal: Cells

    Article Title: Modulation of Protein Synthesis by eIF2α Phosphorylation Protects Cell from Heat Stress-Mediated Apoptosis

    doi: 10.3390/cells7120254

    Figure Lengend Snippet: The IRE1α pathway does not protect cells from heat stress-mediated death. ( a ) Quantitative RT-PCR was performed using total RNA, extracted from Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM. (n = 3 independent experiments). ( b ) Protein expression under conditions of hyperthermia was measured. Following incubation at 42 °C, cell lysates were collected at the indicated times for western blot analysis. ( c ) Cell viability was measured in Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for indicated time periods. ( d ) Caspase 3/7 activity was detected in Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for 12 h. ( e ) Expression of sXBP1 (spliced form) and tXBP (total form) were measured using MEFs treated with 4μ8c at indicated doses in the presence or absence of Tg (300 nM) for 12 h. ( f ) Protein expression under conditions of hyperthermia was measured in the presence or absence of 4μ8c (20 μM). ( g ) Cell viability was measured in Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of 4μ8c (20 μM). * p

    Article Snippet: RNA Extraction and Real-Time PCR Analysis Total RNA was extracted from cells using Trizol (Gene All, Ribo-Ex, Seoul, Korea) and cDNA was synthesized using an iScript cDNA synthesis kit (Bio-Rad, Hercules, CA, USA, BR170-8891).

    Techniques: Quantitative RT-PCR, Incubation, Expressing, Western Blot, Activity Assay