primary antibodies ago2  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc primary antibodies ago2
    Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with <t>AGO2.</t> Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant
    Primary Antibodies Ago2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    1) Product Images from "Negative regulation of lncRNA GAS5 by miR-21"

    Article Title: Negative regulation of lncRNA GAS5 by miR-21

    Journal: Cell Death and Differentiation

    doi: 10.1038/cdd.2013.110

    Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with AGO2. Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant
    Figure Legend Snippet: Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with AGO2. Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Isolation, Quantitative RT-PCR, Immunoprecipitation, Western Blot, Labeling, In Vitro

    ago2 primary antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc ago2 primary antibody
    Ago2 Primary Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ago2 primary antibody/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    primary antibodies against ago2  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc primary antibodies against ago2
    Primary Antibodies Against Ago2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ago2 primary antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc ago2 primary antibody
    Effect of hepatic Argonaute 2 <t>(Ago2)</t> deficiency on glucose metabolism of diabetic obese mice. A, Body weight of liver-specific Ago2-deficiency (L-Ago2) wild-type (WT)- ob/ob (n = 16), and L-Ago2 knockout (KO)- ob/ob (n = 13) mice on normal chow (CD) from age 5 to 12 weeks. B, Food intake normalized by body weight per day of L-Ago2 WT- ob/ob (n = 21), and L-Ago2 KO- ob/ob (n = 24) mice at age 11 weeks. C, Blood glucose levels after 6 hours of fasting of L-Ago2 WT- ob/ob (n = 20) and L-Ago2 KO- ob/ob (n = 16) at age 13 weeks. D, Glucose tolerance test (GTT) and its area under the curve (AUC) analysis in L-Ago2 WT- ob/ob (n = 18) and L-Ago2 KO- ob/ob (n = 18) mice at age 7 weeks. E, Serum insulin levels after 6 hours of fasting of L-Ago2 WT-Lean (n = 3), L-Ago2 WT- ob/ob (n = 4), L-Ago2 KO-Lean (n = 3), and L-Ago2 KO- ob/ob (n = 7) mice at age 13 weeks. F, Insulin tolerance test (ITT) and its AUC analysis in L-Ago2 WT- ob/ob (n = 18) and L-Ago2 KO- ob/ob (n = 18) mice at age 9 weeks. G, Messenger RNA (mRNA) expression levels of de novo lipogenesis and gluconeogenesis in livers of L-Ago2 WT- Lean (n = 5), L-Ago2 WT- ob/ob (n = 10), L-Ago2 KO- Lean (n = 5), and L-Ago2 KO- ob/ob (n = 4) mice after overnight fasting at age 13 weeks. H, Metabolic disease–associated microRNAs (MD-miRNAs) and I, their target mRNA expression levels in livers of L-Ago2 WT- Lean (n = 5), L-Ago2 WT- ob/ob (n = 10), L-Ago2 KO- Lean (n = 5), and L-Ago2 KO- ob/ob (n = 4) mice after overnight fasting at age 13 weeks. Data are shown as mean ± SEM of group size (n). Statistical analyses were performed by unpaired 2-tailed t test for C, D, and F or by 2-way analysis of variance followed by Tukey post hoc test for E, G, H, and I. * P less than or equal to .05, ** P less than or equal to .01, *** P less than or equal to .001.
    Ago2 Primary Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ago2 primary antibody/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ago2 primary antibody - by Bioz Stars, 2023-03
    86/100 stars

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    1) Product Images from "Hepatic Ago2 Regulates PPARα for Oxidative Metabolism Linked to Glycemic Control in Obesity and Post Bariatric Surgery"

    Article Title: Hepatic Ago2 Regulates PPARα for Oxidative Metabolism Linked to Glycemic Control in Obesity and Post Bariatric Surgery

    Journal: Endocrinology

    doi: 10.1210/endocr/bqab007

    Effect of hepatic Argonaute 2 (Ago2) deficiency on glucose metabolism of diabetic obese mice. A, Body weight of liver-specific Ago2-deficiency (L-Ago2) wild-type (WT)- ob/ob (n = 16), and L-Ago2 knockout (KO)- ob/ob (n = 13) mice on normal chow (CD) from age 5 to 12 weeks. B, Food intake normalized by body weight per day of L-Ago2 WT- ob/ob (n = 21), and L-Ago2 KO- ob/ob (n = 24) mice at age 11 weeks. C, Blood glucose levels after 6 hours of fasting of L-Ago2 WT- ob/ob (n = 20) and L-Ago2 KO- ob/ob (n = 16) at age 13 weeks. D, Glucose tolerance test (GTT) and its area under the curve (AUC) analysis in L-Ago2 WT- ob/ob (n = 18) and L-Ago2 KO- ob/ob (n = 18) mice at age 7 weeks. E, Serum insulin levels after 6 hours of fasting of L-Ago2 WT-Lean (n = 3), L-Ago2 WT- ob/ob (n = 4), L-Ago2 KO-Lean (n = 3), and L-Ago2 KO- ob/ob (n = 7) mice at age 13 weeks. F, Insulin tolerance test (ITT) and its AUC analysis in L-Ago2 WT- ob/ob (n = 18) and L-Ago2 KO- ob/ob (n = 18) mice at age 9 weeks. G, Messenger RNA (mRNA) expression levels of de novo lipogenesis and gluconeogenesis in livers of L-Ago2 WT- Lean (n = 5), L-Ago2 WT- ob/ob (n = 10), L-Ago2 KO- Lean (n = 5), and L-Ago2 KO- ob/ob (n = 4) mice after overnight fasting at age 13 weeks. H, Metabolic disease–associated microRNAs (MD-miRNAs) and I, their target mRNA expression levels in livers of L-Ago2 WT- Lean (n = 5), L-Ago2 WT- ob/ob (n = 10), L-Ago2 KO- Lean (n = 5), and L-Ago2 KO- ob/ob (n = 4) mice after overnight fasting at age 13 weeks. Data are shown as mean ± SEM of group size (n). Statistical analyses were performed by unpaired 2-tailed t test for C, D, and F or by 2-way analysis of variance followed by Tukey post hoc test for E, G, H, and I. * P less than or equal to .05, ** P less than or equal to .01, *** P less than or equal to .001.
    Figure Legend Snippet: Effect of hepatic Argonaute 2 (Ago2) deficiency on glucose metabolism of diabetic obese mice. A, Body weight of liver-specific Ago2-deficiency (L-Ago2) wild-type (WT)- ob/ob (n = 16), and L-Ago2 knockout (KO)- ob/ob (n = 13) mice on normal chow (CD) from age 5 to 12 weeks. B, Food intake normalized by body weight per day of L-Ago2 WT- ob/ob (n = 21), and L-Ago2 KO- ob/ob (n = 24) mice at age 11 weeks. C, Blood glucose levels after 6 hours of fasting of L-Ago2 WT- ob/ob (n = 20) and L-Ago2 KO- ob/ob (n = 16) at age 13 weeks. D, Glucose tolerance test (GTT) and its area under the curve (AUC) analysis in L-Ago2 WT- ob/ob (n = 18) and L-Ago2 KO- ob/ob (n = 18) mice at age 7 weeks. E, Serum insulin levels after 6 hours of fasting of L-Ago2 WT-Lean (n = 3), L-Ago2 WT- ob/ob (n = 4), L-Ago2 KO-Lean (n = 3), and L-Ago2 KO- ob/ob (n = 7) mice at age 13 weeks. F, Insulin tolerance test (ITT) and its AUC analysis in L-Ago2 WT- ob/ob (n = 18) and L-Ago2 KO- ob/ob (n = 18) mice at age 9 weeks. G, Messenger RNA (mRNA) expression levels of de novo lipogenesis and gluconeogenesis in livers of L-Ago2 WT- Lean (n = 5), L-Ago2 WT- ob/ob (n = 10), L-Ago2 KO- Lean (n = 5), and L-Ago2 KO- ob/ob (n = 4) mice after overnight fasting at age 13 weeks. H, Metabolic disease–associated microRNAs (MD-miRNAs) and I, their target mRNA expression levels in livers of L-Ago2 WT- Lean (n = 5), L-Ago2 WT- ob/ob (n = 10), L-Ago2 KO- Lean (n = 5), and L-Ago2 KO- ob/ob (n = 4) mice after overnight fasting at age 13 weeks. Data are shown as mean ± SEM of group size (n). Statistical analyses were performed by unpaired 2-tailed t test for C, D, and F or by 2-way analysis of variance followed by Tukey post hoc test for E, G, H, and I. * P less than or equal to .05, ** P less than or equal to .01, *** P less than or equal to .001.

    Techniques Used: Knock-Out, Expressing

    Effect of hepatic Argonaute 2 (Ago2) deficiency on glucose metabolism in a long-term high-fat diet (HFD) condition. A, Body weight of liver-specific Ago2-deficiency (L-Ago2) wild-type (WT) and L-Ago2 knockout (KO) mice on an HFD at age 23 (n = 22 and 25, respectively), 30 (n = 22 and 25, respectively), and 34 (n = 9 and 10, respectively) weeks. B, Blood glucose levels after 6 hours of fasting of L-Ago2 WT (n = 9) and L-Ago2 KO (n = 10) mice at age 34 weeks. C, Serum insulin levels of L-Ago2 WT (n = 9) and L-Ago2 KO (n = 10) mice at age 34 weeks. D, Glucose tolerance test (GTT) and its area under the curve (AUC) analysis in L-Ago2 WT (n = 9) and L-Ago2 KO (n = 10) mice at age 30 weeks. E, Insulin tolerance test (ITT) and its AUC analysis in L-Ago2 WT (n = 9), and L-Ago2 KO (n = 10) mice at age 31 weeks. F, Metabolic disease–associated microRNAs (MD-miRNAs) and their G, target messenger RNA (mRNA) expression levels in livers of L-Ago2 WT (n = 9) and L-Ago2 KO (n = 9) mice at age 34 weeks. Data are shown as mean ± SEM of group size (n). Statistical analyses were performed by ordinary one-way analysis of variance, followed by Tukey post hoc test for A or unpaired 2-tailed test for B to G. * P less than or equal to .05, ** P less than or equal to .01, *** P less than or equal to .001.
    Figure Legend Snippet: Effect of hepatic Argonaute 2 (Ago2) deficiency on glucose metabolism in a long-term high-fat diet (HFD) condition. A, Body weight of liver-specific Ago2-deficiency (L-Ago2) wild-type (WT) and L-Ago2 knockout (KO) mice on an HFD at age 23 (n = 22 and 25, respectively), 30 (n = 22 and 25, respectively), and 34 (n = 9 and 10, respectively) weeks. B, Blood glucose levels after 6 hours of fasting of L-Ago2 WT (n = 9) and L-Ago2 KO (n = 10) mice at age 34 weeks. C, Serum insulin levels of L-Ago2 WT (n = 9) and L-Ago2 KO (n = 10) mice at age 34 weeks. D, Glucose tolerance test (GTT) and its area under the curve (AUC) analysis in L-Ago2 WT (n = 9) and L-Ago2 KO (n = 10) mice at age 30 weeks. E, Insulin tolerance test (ITT) and its AUC analysis in L-Ago2 WT (n = 9), and L-Ago2 KO (n = 10) mice at age 31 weeks. F, Metabolic disease–associated microRNAs (MD-miRNAs) and their G, target messenger RNA (mRNA) expression levels in livers of L-Ago2 WT (n = 9) and L-Ago2 KO (n = 9) mice at age 34 weeks. Data are shown as mean ± SEM of group size (n). Statistical analyses were performed by ordinary one-way analysis of variance, followed by Tukey post hoc test for A or unpaired 2-tailed test for B to G. * P less than or equal to .05, ** P less than or equal to .01, *** P less than or equal to .001.

    Techniques Used: Knock-Out, Expressing

    Effects of vertical sleeve gastrectomy (VSG) surgery on glycemic control and metabolic disease–associated microRNA (MD-miRNA) expression in liver-specific Ago2-deficiency (L-Ago2) knockout (KO) mice. A, Body weight gain of L-Ago2 wild-type (WT)- SHAM (n = 7) and L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 6) mice post surgery. B, Body weight loss of L-Ago2 WT- SHAM (n = 7) and L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 5) mice pre surgery and post surgery. C, Blood glucose levels after 6 hours of fasting of L-Ago2 WT- SHAM (n = 7), L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 6) mice 4 weeks post surgery. D, Serum insulin levels after 6 hours of fasting of L-Ago2 WT- SHAM (n = 7), L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 6) mice 4 weeks post surgery. E, hematoxylin-eosin–stained liver section of L-Ago2 WT- SHAM , L-Ago2 WT- VSG , L-Ago2 KO- SHAM , and L-Ago2 KO- VSG mice 4 weeks post surgery. F, Plasma alanine aminotransferase (ALT) levels of L-Ago2 WT- SHAM (n = 4), L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 6), and L-Ago2 KO- VSG (n = 6) mice 4 weeks post surgery. G, MD-miRNAs and their target messenger RNA (mRNA), de novo lipogenesis, and H, gluconeogenesis gene expression levels in livers of L-Ago2 WT- SHAM (n = 6), L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 6) mice 4 weeks post surgery. Data are shown as mean ± SEM of group size (n). Statistical analyses were performed by multiple t test for A or by unpaired 2-tailed test for C and F, or by 2-way analysis of variance followed by Tukey post hoc test for B, G, and H. * P less than or equal to .05, * P less than or equal to .01, *** P less than or equal to .001.
    Figure Legend Snippet: Effects of vertical sleeve gastrectomy (VSG) surgery on glycemic control and metabolic disease–associated microRNA (MD-miRNA) expression in liver-specific Ago2-deficiency (L-Ago2) knockout (KO) mice. A, Body weight gain of L-Ago2 wild-type (WT)- SHAM (n = 7) and L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 6) mice post surgery. B, Body weight loss of L-Ago2 WT- SHAM (n = 7) and L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 5) mice pre surgery and post surgery. C, Blood glucose levels after 6 hours of fasting of L-Ago2 WT- SHAM (n = 7), L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 6) mice 4 weeks post surgery. D, Serum insulin levels after 6 hours of fasting of L-Ago2 WT- SHAM (n = 7), L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 6) mice 4 weeks post surgery. E, hematoxylin-eosin–stained liver section of L-Ago2 WT- SHAM , L-Ago2 WT- VSG , L-Ago2 KO- SHAM , and L-Ago2 KO- VSG mice 4 weeks post surgery. F, Plasma alanine aminotransferase (ALT) levels of L-Ago2 WT- SHAM (n = 4), L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 6), and L-Ago2 KO- VSG (n = 6) mice 4 weeks post surgery. G, MD-miRNAs and their target messenger RNA (mRNA), de novo lipogenesis, and H, gluconeogenesis gene expression levels in livers of L-Ago2 WT- SHAM (n = 6), L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 6) mice 4 weeks post surgery. Data are shown as mean ± SEM of group size (n). Statistical analyses were performed by multiple t test for A or by unpaired 2-tailed test for C and F, or by 2-way analysis of variance followed by Tukey post hoc test for B, G, and H. * P less than or equal to .05, * P less than or equal to .01, *** P less than or equal to .001.

    Techniques Used: Expressing, Knock-Out, Staining

    Hepatic Argonaute 2 (Ago2) deficiency enhances the expression of the peroxisome proliferator–activated receptor α (PPARα) pathway. A, Messenger RNA (mRNA) expression of Tfam -mitochondrial genes in livers of liver-specific Ago2-deficiency (L-Ago2) wild-type (WT) (n = 9) and L-Ago2 knockout (KO) (n = 9) mice at age 34 weeks. B, Western blot analysis of PPARα protein expression in the liver of L-Ago2 WT (n = 6) and L-Ago2 KO (n = 6) mice at age 34 weeks. C, miR-27a and miR-27b expression in livers of L-Ago2 WT (n = 9) and L-Ago2 KO (n = 9) mice at age 34 weeks. D, PPARα and its target mRNA expression in livers of L-Ago2 WT (n = 9) and L-Ago2 KO (n = 9) mice at age 34 weeks. E, Western blot analysis of PPARα protein expression in the liver of L-Ago2 WT- SHAM (n = 3), L-Ago2 WT- VSG (n = 3), L-Ago2 KO- SHAM (n = 3), and L-Ago2 KO- VSG (n = 3) mice 4 weeks post surgery. F, miR-27a and miR-27b expression in livers of L-Ago2 WT- SHAM (n = 6), L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 6) mice 4 weeks post surgery. G, PPARα and its target mRNA expression in livers of L-Ago2 WT- SHAM (n = 6), L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 6) mice 4 weeks post surgery. NS, nonspecific band. Data are shown as mean ± SEM of group size (n). Statistical analyses were performed by unpaired 2-tailed test for A to D, by multiple t test for E, or by 2-way analysis of variance followed by Tukey post hoc test for F and G. * P less than or equal to .05, * P less than or equal to .01, *** P less than or equal to .001.
    Figure Legend Snippet: Hepatic Argonaute 2 (Ago2) deficiency enhances the expression of the peroxisome proliferator–activated receptor α (PPARα) pathway. A, Messenger RNA (mRNA) expression of Tfam -mitochondrial genes in livers of liver-specific Ago2-deficiency (L-Ago2) wild-type (WT) (n = 9) and L-Ago2 knockout (KO) (n = 9) mice at age 34 weeks. B, Western blot analysis of PPARα protein expression in the liver of L-Ago2 WT (n = 6) and L-Ago2 KO (n = 6) mice at age 34 weeks. C, miR-27a and miR-27b expression in livers of L-Ago2 WT (n = 9) and L-Ago2 KO (n = 9) mice at age 34 weeks. D, PPARα and its target mRNA expression in livers of L-Ago2 WT (n = 9) and L-Ago2 KO (n = 9) mice at age 34 weeks. E, Western blot analysis of PPARα protein expression in the liver of L-Ago2 WT- SHAM (n = 3), L-Ago2 WT- VSG (n = 3), L-Ago2 KO- SHAM (n = 3), and L-Ago2 KO- VSG (n = 3) mice 4 weeks post surgery. F, miR-27a and miR-27b expression in livers of L-Ago2 WT- SHAM (n = 6), L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 6) mice 4 weeks post surgery. G, PPARα and its target mRNA expression in livers of L-Ago2 WT- SHAM (n = 6), L-Ago2 WT- VSG (n = 6), L-Ago2 KO- SHAM (n = 7), and L-Ago2 KO- VSG (n = 6) mice 4 weeks post surgery. NS, nonspecific band. Data are shown as mean ± SEM of group size (n). Statistical analyses were performed by unpaired 2-tailed test for A to D, by multiple t test for E, or by 2-way analysis of variance followed by Tukey post hoc test for F and G. * P less than or equal to .05, * P less than or equal to .01, *** P less than or equal to .001.

    Techniques Used: Expressing, Knock-Out, Western Blot

    Effects of peroxisome proliferator-activated receptor (PPAR) agonist in Argonaute 2 (Ago2)-deficient primary hepatocytes. A, messenger RNA (mRNA) expression of Tfam -mitochondrial genes in liver-specific Ago2-deficiency (L-Ago2) wild-type (WT) (n = 6), and L-Ago2 knockout (KO) (n = 6) primary hepatocytes treated with or without 10-µM WY14643 (a PPARα agonist) for 24 hours. B, miR-27a and miR-27b expression in L-Ago2 WT (n = 6) and L-Ago2 KO (n = 6) primary hepatocytes treated with or without 10-µM WY14643 (PPARα agonist) for 24 hours. C, Oxygen consumption rate (OCR) of L-Ago2 WT (n = 12) and KO (n = 10–12) primary hepatocytes were measured in the presence or absence of a PPARα agonist (WY14643: 10 μM) pretreatment for 48 hours. D, PPARα target mRNA expression in L-Ago2 WT (n = 6) and L-Ago2 KO (n = 6) primary hepatocytes treated with or without 10-µM WY14643 (PPARα agonist) for 24 hours. Data are shown as mean± SEM. Statistical analyses were performed by 2-way analysis of variance for A, B, and D; and ordinary 1-way analysis of variance followed by Tukey post hoc test for C. * P less than or equal to .05, * P less than or equal to .01, *** P less than or equal to .001.
    Figure Legend Snippet: Effects of peroxisome proliferator-activated receptor (PPAR) agonist in Argonaute 2 (Ago2)-deficient primary hepatocytes. A, messenger RNA (mRNA) expression of Tfam -mitochondrial genes in liver-specific Ago2-deficiency (L-Ago2) wild-type (WT) (n = 6), and L-Ago2 knockout (KO) (n = 6) primary hepatocytes treated with or without 10-µM WY14643 (a PPARα agonist) for 24 hours. B, miR-27a and miR-27b expression in L-Ago2 WT (n = 6) and L-Ago2 KO (n = 6) primary hepatocytes treated with or without 10-µM WY14643 (PPARα agonist) for 24 hours. C, Oxygen consumption rate (OCR) of L-Ago2 WT (n = 12) and KO (n = 10–12) primary hepatocytes were measured in the presence or absence of a PPARα agonist (WY14643: 10 μM) pretreatment for 48 hours. D, PPARα target mRNA expression in L-Ago2 WT (n = 6) and L-Ago2 KO (n = 6) primary hepatocytes treated with or without 10-µM WY14643 (PPARα agonist) for 24 hours. Data are shown as mean± SEM. Statistical analyses were performed by 2-way analysis of variance for A, B, and D; and ordinary 1-way analysis of variance followed by Tukey post hoc test for C. * P less than or equal to .05, * P less than or equal to .01, *** P less than or equal to .001.

    Techniques Used: Expressing, Knock-Out

    A proposed molecular mechanism of the role of hepatic Argonaute 2 (Ago2) in obesity, post bariatric surgery, and in peroxisome proliferator–activated receptor α (PPARα) response. Hepatic Ago2 is required for the expression of metabolic disease–associated microRNA (MD-miRNA), which is enhanced in obesity but suppressed post bariatric surgery. Hepatic Ago2-dependent RNA silencing plays a critical role in gene regulation relevant to mitochondrial functions, including PPARα, adenosine 5′-monophosphate–activated protein kinase α (AMPKα1), and PPAR γ coactivator (PGC1α), and PPARα-mediated oxidative metabolism in obesity, post–vertical sleeve gastrectomy (VSG), and PPARα response.
    Figure Legend Snippet: A proposed molecular mechanism of the role of hepatic Argonaute 2 (Ago2) in obesity, post bariatric surgery, and in peroxisome proliferator–activated receptor α (PPARα) response. Hepatic Ago2 is required for the expression of metabolic disease–associated microRNA (MD-miRNA), which is enhanced in obesity but suppressed post bariatric surgery. Hepatic Ago2-dependent RNA silencing plays a critical role in gene regulation relevant to mitochondrial functions, including PPARα, adenosine 5′-monophosphate–activated protein kinase α (AMPKα1), and PPAR γ coactivator (PGC1α), and PPARα-mediated oxidative metabolism in obesity, post–vertical sleeve gastrectomy (VSG), and PPARα response.

    Techniques Used: Expressing

    ago2 primary antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc ago2 primary antibody
    A) Schematic presentation of the vector composition for miR-183/96/182 inducible inhibition using the TuD approach. B) The morphology of transduced hiPSCs expressing T uD for inhibition of the miR-183/96/182 cluster or SCR control, as determined by bright field and fluorescent microscopy. C) Quantification of cells expressing mCherry, as determined by flow cytometry. D) Quantification of TuD miR-183/96/182 transcript levels in <t>Ago2-immunoprecipitated</t> (Ago2-IP) fraction, as demonstrated by RT-qPCR. Data are depicted as mean + SD of triplicates. E) Western blot analysis of Ago2 levels in Ago2-IP reaction. Lower part: amidoblack-stained PVDF membrane represents a loading control.
    Ago2 Primary Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "miR-183/96/182 cluster is an important morphogenetic factor targeting PAX6 expression in differentiating human retinal organoids"

    Article Title: miR-183/96/182 cluster is an important morphogenetic factor targeting PAX6 expression in differentiating human retinal organoids

    Journal: bioRxiv

    doi: 10.1101/2020.04.01.019539

    A) Schematic presentation of the vector composition for miR-183/96/182 inducible inhibition using the TuD approach. B) The morphology of transduced hiPSCs expressing T uD for inhibition of the miR-183/96/182 cluster or SCR control, as determined by bright field and fluorescent microscopy. C) Quantification of cells expressing mCherry, as determined by flow cytometry. D) Quantification of TuD miR-183/96/182 transcript levels in Ago2-immunoprecipitated (Ago2-IP) fraction, as demonstrated by RT-qPCR. Data are depicted as mean + SD of triplicates. E) Western blot analysis of Ago2 levels in Ago2-IP reaction. Lower part: amidoblack-stained PVDF membrane represents a loading control.
    Figure Legend Snippet: A) Schematic presentation of the vector composition for miR-183/96/182 inducible inhibition using the TuD approach. B) The morphology of transduced hiPSCs expressing T uD for inhibition of the miR-183/96/182 cluster or SCR control, as determined by bright field and fluorescent microscopy. C) Quantification of cells expressing mCherry, as determined by flow cytometry. D) Quantification of TuD miR-183/96/182 transcript levels in Ago2-immunoprecipitated (Ago2-IP) fraction, as demonstrated by RT-qPCR. Data are depicted as mean + SD of triplicates. E) Western blot analysis of Ago2 levels in Ago2-IP reaction. Lower part: amidoblack-stained PVDF membrane represents a loading control.

    Techniques Used: Plasmid Preparation, Inhibition, Expressing, Microscopy, Flow Cytometry, Immunoprecipitation, Quantitative RT-PCR, Western Blot, Staining

    ago2 primary antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc ago2 primary antibody
    ( a ) Cell morphology and GFP expression upon cumate-induced expression of miR-145 sponge, as observed by light and fluorescent microscopy. Scale bars represent 100 μm. ( b ) Quantification of sponge transcript levels in <t>Ago2-immunoprecipitated</t> (Ago2 IP) and control IgG (IgG IP) fractions, as demonstrated by RT-qPCR. Data are depicted as mean + SD of triplicates. The length of the transcript was checked by standard agarose electrophoresis (graph inset). Full-length gel is presented in . ( c ) Western blot analysis of Ago2 levels in Ago2 IP and control IgG IP fractions. Full-length blot is presented in . ( d ) Expression of c-Myc and Sox2 upon miR-145 sponge induction, as determined by western blot analysis. Experiments were performed on three different HEK293T clones expressing miR-145 sponge or empty vector. One representative western blot is shown. Full-length blot is presented in . Quantification of western blots is provided under each western blot. Data represents fold change relative to an empty vector control without cumate ± SD. β-Actin was used as a loading control. ( e ) Levels of MYC and SOX2 transcripts associated with Ago2 upon miR-145 sponge expression. Data are depicted as mean ± SD of triplicates.
    Ago2 Primary Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "miRNAsong: a web-based tool for generation and testing of miRNA sponge constructs in silico"

    Article Title: miRNAsong: a web-based tool for generation and testing of miRNA sponge constructs in silico

    Journal: Scientific Reports

    doi: 10.1038/srep36625

    ( a ) Cell morphology and GFP expression upon cumate-induced expression of miR-145 sponge, as observed by light and fluorescent microscopy. Scale bars represent 100 μm. ( b ) Quantification of sponge transcript levels in Ago2-immunoprecipitated (Ago2 IP) and control IgG (IgG IP) fractions, as demonstrated by RT-qPCR. Data are depicted as mean + SD of triplicates. The length of the transcript was checked by standard agarose electrophoresis (graph inset). Full-length gel is presented in . ( c ) Western blot analysis of Ago2 levels in Ago2 IP and control IgG IP fractions. Full-length blot is presented in . ( d ) Expression of c-Myc and Sox2 upon miR-145 sponge induction, as determined by western blot analysis. Experiments were performed on three different HEK293T clones expressing miR-145 sponge or empty vector. One representative western blot is shown. Full-length blot is presented in . Quantification of western blots is provided under each western blot. Data represents fold change relative to an empty vector control without cumate ± SD. β-Actin was used as a loading control. ( e ) Levels of MYC and SOX2 transcripts associated with Ago2 upon miR-145 sponge expression. Data are depicted as mean ± SD of triplicates.
    Figure Legend Snippet: ( a ) Cell morphology and GFP expression upon cumate-induced expression of miR-145 sponge, as observed by light and fluorescent microscopy. Scale bars represent 100 μm. ( b ) Quantification of sponge transcript levels in Ago2-immunoprecipitated (Ago2 IP) and control IgG (IgG IP) fractions, as demonstrated by RT-qPCR. Data are depicted as mean + SD of triplicates. The length of the transcript was checked by standard agarose electrophoresis (graph inset). Full-length gel is presented in . ( c ) Western blot analysis of Ago2 levels in Ago2 IP and control IgG IP fractions. Full-length blot is presented in . ( d ) Expression of c-Myc and Sox2 upon miR-145 sponge induction, as determined by western blot analysis. Experiments were performed on three different HEK293T clones expressing miR-145 sponge or empty vector. One representative western blot is shown. Full-length blot is presented in . Quantification of western blots is provided under each western blot. Data represents fold change relative to an empty vector control without cumate ± SD. β-Actin was used as a loading control. ( e ) Levels of MYC and SOX2 transcripts associated with Ago2 upon miR-145 sponge expression. Data are depicted as mean ± SD of triplicates.

    Techniques Used: Expressing, Microscopy, Immunoprecipitation, Quantitative RT-PCR, Electrophoresis, Western Blot, Clone Assay, Plasmid Preparation

    primary antibodies ago2  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc primary antibodies ago2
    Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with <t>AGO2.</t> Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant
    Primary Antibodies Ago2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Negative regulation of lncRNA GAS5 by miR-21"

    Article Title: Negative regulation of lncRNA GAS5 by miR-21

    Journal: Cell Death and Differentiation

    doi: 10.1038/cdd.2013.110

    Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with AGO2. Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant
    Figure Legend Snippet: Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with AGO2. Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Isolation, Quantitative RT-PCR, Immunoprecipitation, Western Blot, Labeling, In Vitro

    primary antibodies ago2  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc primary antibodies ago2
    Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with <t>AGO2.</t> Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant
    Primary Antibodies Ago2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Negative regulation of lncRNA GAS5 by miR-21"

    Article Title: Negative regulation of lncRNA GAS5 by miR-21

    Journal: Cell Death and Differentiation

    doi: 10.1038/cdd.2013.110

    Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with AGO2. Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant
    Figure Legend Snippet: Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with AGO2. Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Isolation, Quantitative RT-PCR, Immunoprecipitation, Western Blot, Labeling, In Vitro

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    Cell Signaling Technology Inc primary antibodies ago2
    Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with <t>AGO2.</t> Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant
    Primary Antibodies Ago2, supplied by Cell Signaling Technology Inc, 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/primary antibodies ago2/product/Cell Signaling Technology Inc
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    ago2 primary antibody  (Cell Signaling Technology Inc)
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    Cell Signaling Technology Inc ago2 primary antibody
    Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with <t>AGO2.</t> Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant
    Ago2 Primary Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    primary antibodies against ago2  (Cell Signaling Technology Inc)
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    Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with <t>AGO2.</t> Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant
    Primary Antibodies Against Ago2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary antibodies against ago2/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    primary antibodies against ago2 - by Bioz Stars, 2023-03
    86/100 stars
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    Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with AGO2. Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant

    Journal: Cell Death and Differentiation

    Article Title: Negative regulation of lncRNA GAS5 by miR-21

    doi: 10.1038/cdd.2013.110

    Figure Lengend Snippet: Post-transcriptional regulation of GAS5 and miR-21. (a) Whereas anti-miR-21 upregulates GAS5, it has no effect on U44 and U77, which are imbedded in the GAS5 gene. MCF-7 cells were transfected with scrambled oligo or anti-miR-21 and the rest of the procedure was same as in Figure 1c. Primers for U44 and U77 were U44-RT-5.1 and U44-RT-3.1; U77-RT-5.1 and U77-RT-3.1, respectively. (b and c) Effect of ectopic expression of GAS5 or GAS5–siRNA on pri-miR-21, pre-miR-21 and mature miR-21. MCF-7 cells were transfected with vector alone or GAS5, or control siRNA or GAS5–siRNA. Total RNA was isolated from the cells 24 h later, followed by qRT-PCR. Primers for pri-miR-21 were Pri-miR-21-RT-5.1 and Pri-miR-21-RT-3.1; Pre-miR-21-RT-5.1 and Pre-miR-21-RT-3.1. (d) Association of GAS5 and miR-21 with AGO2. Cellular lysates from MCF-7 cells were used for RNA immunoprecipitation with AGO2 antibody. Detection of AGO2 using IP-western (left panel), and detection of GAS5 and miR-21 using qRT-PCR (right panel). (e and f) Identification of GAS5 and miR-21 in the same RISC complex by RNA precipitation. (e) A procedure for making biotin-labeled RNA probes and RNA precipitation. In vitro transcribed RNA probes were made using T7 RNA polymerase, followed by precipitation assays as described in the Materials and Methods section. (f) Detection of miR-21 in the GAS5 RNA-precipitated samples. Error bars represent S.E.M., n=3. *P<0.05; **P<0.01; n.s., not significant

    Article Snippet: Primary antibodies Ago2 and PTEN were purchased from Cell Signaling (Danvers, MA, USA); PDCD4 from Epitomics (Burlingame, CA, USA); GAPDH from Protein Tech (Chicago, IL, USA) and β -actin from Sigma (St. Louis, MO, USA).

    Techniques: Transfection, Expressing, Plasmid Preparation, Isolation, Quantitative RT-PCR, Immunoprecipitation, Western Blot, Labeling, In Vitro