cdna synthesis cdna  (Qiagen)

 
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    Name:
    miScript II RT Kit
    Description:
    For reverse transcription of total RNA containing miRNA using the miScript PCR System Kit contents Qiagen miScript II RT Kit 12 rxns SYBR Green based Single step cDNA Synthesis Quantification of miRNA and mRNA For Reverse Transcription of Total RNA Containing miRNA Using the miScript PCR System Includes miScript Reverse Transcriptase Mix 10x miScript Nucleics Mix 5x miScript HiSpec Buffer 5x miScript HiFlex Buffer RNase free Water Benefits cDNA for sensitive and specific miRNA detection A single cDNA enables quantification of multiple miRNAs Quantification of miRNA and mRNA from the same cDNA Quantification of mature and precursor miRNA from the same cDNA Proprietary synthetic RNA to assess reverse transcription efficiency
    Catalog Number:
    218160
    Price:
    133
    Category:
    miScript II RT Kit
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    Structured Review

    Qiagen cdna synthesis cdna
    miScript II RT Kit
    For reverse transcription of total RNA containing miRNA using the miScript PCR System Kit contents Qiagen miScript II RT Kit 12 rxns SYBR Green based Single step cDNA Synthesis Quantification of miRNA and mRNA For Reverse Transcription of Total RNA Containing miRNA Using the miScript PCR System Includes miScript Reverse Transcriptase Mix 10x miScript Nucleics Mix 5x miScript HiSpec Buffer 5x miScript HiFlex Buffer RNase free Water Benefits cDNA for sensitive and specific miRNA detection A single cDNA enables quantification of multiple miRNAs Quantification of miRNA and mRNA from the same cDNA Quantification of mature and precursor miRNA from the same cDNA Proprietary synthetic RNA to assess reverse transcription efficiency
    https://www.bioz.com/result/cdna synthesis cdna/product/Qiagen
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    cdna synthesis cdna - by Bioz Stars, 2020-08
    91/100 stars

    Images

    1) Product Images from "Proof-of-concept study: profile of circulating microRNAs in Bovine serum harvested during acute and persistent FMDV infection"

    Article Title: Proof-of-concept study: profile of circulating microRNAs in Bovine serum harvested during acute and persistent FMDV infection

    Journal: Virology Journal

    doi: 10.1186/s12985-017-0743-3

    Schematic overview of miRNA profiling study. a Serum was collected from three different groups of FMDV-infected cattle: acutely infected (viremic; 3–4 dpi), persistently infected (“FMDV carriers”; 35 dpi) and convalescent (“non-carriers”; 35 dpi), and were compared to uninfected controls. Each group comprised serum samples from three animals. b miRNAs were purified from individual serum samples. The purified miRNAs were reverse transcribed into complementary DNA (cDNA). The cDNA samples were then analyzed by RT-PCR on bovine miRNome miRNA array plates containing primers to 169 different bovine miRNAs. The results obtained indicated which miRNAs were left unchanged, up-regulated, or down-regulated in response to FMDV infection. c schematic of the organization of the miRNA PCR array plates utilized in this study
    Figure Legend Snippet: Schematic overview of miRNA profiling study. a Serum was collected from three different groups of FMDV-infected cattle: acutely infected (viremic; 3–4 dpi), persistently infected (“FMDV carriers”; 35 dpi) and convalescent (“non-carriers”; 35 dpi), and were compared to uninfected controls. Each group comprised serum samples from three animals. b miRNAs were purified from individual serum samples. The purified miRNAs were reverse transcribed into complementary DNA (cDNA). The cDNA samples were then analyzed by RT-PCR on bovine miRNome miRNA array plates containing primers to 169 different bovine miRNAs. The results obtained indicated which miRNAs were left unchanged, up-regulated, or down-regulated in response to FMDV infection. c schematic of the organization of the miRNA PCR array plates utilized in this study

    Techniques Used: Infection, Purification, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction

    2) Product Images from "miR-125a regulates cell cycle, proliferation, and apoptosis by targeting the ErbB Pathway in Acute Myeloid Leukemia"

    Article Title: miR-125a regulates cell cycle, proliferation, and apoptosis by targeting the ErbB Pathway in Acute Myeloid Leukemia

    Journal: Leukemia research

    doi: 10.1016/j.leukres.2013.12.021

    miR-125a is epigenetically silenced in AML
    Figure Legend Snippet: miR-125a is epigenetically silenced in AML

    Techniques Used:

    Restored miR-125a expression causes decreased cell proliferation, cell cycle progression, and enhanced apoptosis
    Figure Legend Snippet: Restored miR-125a expression causes decreased cell proliferation, cell cycle progression, and enhanced apoptosis

    Techniques Used: Expressing

    3) Product Images from "BAF53b, a Neuron-Specific Nucleosome Remodeling Factor, Is Induced after Learning and Facilitates Long-Term Memory Consolidation"

    Article Title: BAF53b, a Neuron-Specific Nucleosome Remodeling Factor, Is Induced after Learning and Facilitates Long-Term Memory Consolidation

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.3220-16.2017

    BAF53b function is necessary in the LA for the consolidation of auditory fear memory during long-term memory formation. A , Top, Construct map for AAV-nontarget shRNA and AAV- Baf53b shRNA. Bottom, Representative AAV-infected neurons (red) in the LA. Nuclei (blue) were stained with DAPI. Scale bar, 200 μm. B , Western blot showing BAF53b protein levels in the LA of naive mice ( n = 3), nontarget shRNA-expressing mice ( n = 3), and Baf53b shRNA-expressing mice ( n = 3). Top, Representative Western blot. Bottom, BAF53b expression histograms. The y -axis indicates the BAF53b protein level for each condition relative to naive control. The BAF53b protein level for each condition was normalized to the GAPDH level. n values indicate the number of trials. C , Western blot showing protein levels of BAF53a (left, anti-BAF53a) or BAF53b (right, anti-BAF53b) in HEK293T cells infected with either HSV-BAF53a or HSV-BAF53b. No infection condition was included as a control. GAPDH was used as a loading control. D , Top, Western blot showing BAF53b protein levels in the LA of Baf53b +/− (HET, n = 3) and WT ( n = 3) mice. Bottom, BAF53b expression histograms. The y -axis indicates the BAF53b protein level relative to the WT control. The BAF53b protein level for each condition was normalized to the GAPDH protein level. n values indicate the number of trials. E , Baf53b mRNA levels from LAs expressing either the nontarget shRNA ( n = 3) or the Baf53b shRNA ( n = 2) as measured by qPCR. The y -axis indicates the Baf53b mRNA level relative to the nontarget shRNA control. The Baf53b mRNA level for each condition was normalized to Gapdh mRNA level. F , Top, Experimental procedure. HEK293T cells expressing the nontarget shRNA or Baf53b shRNA were infected with either HSV-BAF53a or HSV-BAF53b ( n =3 for each condition). Middle, Representative Western blot. Bottom, BAF53a and BAF53b expression histograms. The y -axis indicates the BAF53a or BAF53b protein level relative to nontarget shRNA control. The BAF53a or BAF53b protein level for each condition was normalized to the GAPDH level. n values indicate the number of trials. G , Behavioral procedure for auditory fear conditioning with strong shock (0.5 mA). H , Top, Behavioral procedure. Bottom, Tone-induced freezing during retention test. Long-term memory tested 24 h after training in mice injected with either AAV-nontarget shRNA (control, n = 9) or AAV- Baf53b shRNA ( Baf53b KD, n = 15). n values indicate the number of mice. I , Top, Behavioral procedure. Bottom, Tone-induced freezing during retention test. Short-term memory tested 1 h after training (control, n = 11; Baf53b KD, n = 12). n values indicate the number of mice. J–L , Open field test with control ( n = 11) or Baf53b KD ( n = 10) mice. n values indicate the number of mice. J , Representative activity pattern tracked by EthoVision XT (Noldus). K , L , Center-crossings ( K ) and total distance moved ( L ) during 20 min. M , Construct maps for the AAV vectors expressing nontarget shRNA as a control and both knockdown-resistant BAF53b (rBAF53b) and Baf53b shRNA. Red represents mutation sites of rBAF53b. N , BAF53b expression levels from LA of noninfected mice (naive, n = 3), LAs infected with AAV-nontarget shRNA (nontarget shRNA, n = 3), AAV- Baf53b shRNA ( Baf53b shRNA, n = 3), or AAV-rBAF53b- Baf53b shRNA (rBAF53b, n = 3) as measured by Western blot. Top, Representative Western blot. Bottom, BAF53b expression histograms. The y -axis indicates the BAF53b protein level relative to the naive control. The BAF53b protein level for each condition was normalized to the GAPDH protein level. n values indicate the number of trials. O , Top, Behavioral procedure. Bottom, Freezing level measured in mice injected with either the AAV-nontarget shRNA (control, n = 6) or the AAV-rBAF53b- Baf53b shRNA ( n = 6) during long-term fear memory test. n values indicate the number of mice. Data are mean ± SEM. n.s., Not significant ( p > 0.05). * p
    Figure Legend Snippet: BAF53b function is necessary in the LA for the consolidation of auditory fear memory during long-term memory formation. A , Top, Construct map for AAV-nontarget shRNA and AAV- Baf53b shRNA. Bottom, Representative AAV-infected neurons (red) in the LA. Nuclei (blue) were stained with DAPI. Scale bar, 200 μm. B , Western blot showing BAF53b protein levels in the LA of naive mice ( n = 3), nontarget shRNA-expressing mice ( n = 3), and Baf53b shRNA-expressing mice ( n = 3). Top, Representative Western blot. Bottom, BAF53b expression histograms. The y -axis indicates the BAF53b protein level for each condition relative to naive control. The BAF53b protein level for each condition was normalized to the GAPDH level. n values indicate the number of trials. C , Western blot showing protein levels of BAF53a (left, anti-BAF53a) or BAF53b (right, anti-BAF53b) in HEK293T cells infected with either HSV-BAF53a or HSV-BAF53b. No infection condition was included as a control. GAPDH was used as a loading control. D , Top, Western blot showing BAF53b protein levels in the LA of Baf53b +/− (HET, n = 3) and WT ( n = 3) mice. Bottom, BAF53b expression histograms. The y -axis indicates the BAF53b protein level relative to the WT control. The BAF53b protein level for each condition was normalized to the GAPDH protein level. n values indicate the number of trials. E , Baf53b mRNA levels from LAs expressing either the nontarget shRNA ( n = 3) or the Baf53b shRNA ( n = 2) as measured by qPCR. The y -axis indicates the Baf53b mRNA level relative to the nontarget shRNA control. The Baf53b mRNA level for each condition was normalized to Gapdh mRNA level. F , Top, Experimental procedure. HEK293T cells expressing the nontarget shRNA or Baf53b shRNA were infected with either HSV-BAF53a or HSV-BAF53b ( n =3 for each condition). Middle, Representative Western blot. Bottom, BAF53a and BAF53b expression histograms. The y -axis indicates the BAF53a or BAF53b protein level relative to nontarget shRNA control. The BAF53a or BAF53b protein level for each condition was normalized to the GAPDH level. n values indicate the number of trials. G , Behavioral procedure for auditory fear conditioning with strong shock (0.5 mA). H , Top, Behavioral procedure. Bottom, Tone-induced freezing during retention test. Long-term memory tested 24 h after training in mice injected with either AAV-nontarget shRNA (control, n = 9) or AAV- Baf53b shRNA ( Baf53b KD, n = 15). n values indicate the number of mice. I , Top, Behavioral procedure. Bottom, Tone-induced freezing during retention test. Short-term memory tested 1 h after training (control, n = 11; Baf53b KD, n = 12). n values indicate the number of mice. J–L , Open field test with control ( n = 11) or Baf53b KD ( n = 10) mice. n values indicate the number of mice. J , Representative activity pattern tracked by EthoVision XT (Noldus). K , L , Center-crossings ( K ) and total distance moved ( L ) during 20 min. M , Construct maps for the AAV vectors expressing nontarget shRNA as a control and both knockdown-resistant BAF53b (rBAF53b) and Baf53b shRNA. Red represents mutation sites of rBAF53b. N , BAF53b expression levels from LA of noninfected mice (naive, n = 3), LAs infected with AAV-nontarget shRNA (nontarget shRNA, n = 3), AAV- Baf53b shRNA ( Baf53b shRNA, n = 3), or AAV-rBAF53b- Baf53b shRNA (rBAF53b, n = 3) as measured by Western blot. Top, Representative Western blot. Bottom, BAF53b expression histograms. The y -axis indicates the BAF53b protein level relative to the naive control. The BAF53b protein level for each condition was normalized to the GAPDH protein level. n values indicate the number of trials. O , Top, Behavioral procedure. Bottom, Freezing level measured in mice injected with either the AAV-nontarget shRNA (control, n = 6) or the AAV-rBAF53b- Baf53b shRNA ( n = 6) during long-term fear memory test. n values indicate the number of mice. Data are mean ± SEM. n.s., Not significant ( p > 0.05). * p

    Techniques Used: Construct, shRNA, Infection, Staining, Western Blot, Mouse Assay, Expressing, Real-time Polymerase Chain Reaction, Injection, Activity Assay, Mutagenesis

    4) Product Images from "BAF53b, a Neuron-Specific Nucleosome Remodeling Factor, Is Induced after Learning and Facilitates Long-Term Memory Consolidation"

    Article Title: BAF53b, a Neuron-Specific Nucleosome Remodeling Factor, Is Induced after Learning and Facilitates Long-Term Memory Consolidation

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.3220-16.2017

    BAF53b function is necessary in the LA for the consolidation of auditory fear memory during long-term memory formation. A , Top, Construct map for AAV-nontarget shRNA and AAV- Baf53b shRNA. Bottom, Representative AAV-infected neurons (red) in the LA. Nuclei (blue) were stained with DAPI. Scale bar, 200 μm. B , Western blot showing BAF53b protein levels in the LA of naive mice ( n = 3), nontarget shRNA-expressing mice ( n = 3), and Baf53b shRNA-expressing mice ( n = 3). Top, Representative Western blot. Bottom, BAF53b expression histograms. The y -axis indicates the BAF53b protein level for each condition relative to naive control. The BAF53b protein level for each condition was normalized to the GAPDH level. n values indicate the number of trials. C , Western blot showing protein levels of BAF53a (left, anti-BAF53a) or BAF53b (right, anti-BAF53b) in HEK293T cells infected with either HSV-BAF53a or HSV-BAF53b. No infection condition was included as a control. GAPDH was used as a loading control. D , Top, Western blot showing BAF53b protein levels in the LA of Baf53b +/− (HET, n = 3) and WT ( n = 3) mice. Bottom, BAF53b expression histograms. The y -axis indicates the BAF53b protein level relative to the WT control. The BAF53b protein level for each condition was normalized to the GAPDH protein level. n values indicate the number of trials. E , Baf53b mRNA levels from LAs expressing either the nontarget shRNA ( n = 3) or the Baf53b shRNA ( n = 2) as measured by qPCR. The y -axis indicates the Baf53b mRNA level relative to the nontarget shRNA control. The Baf53b mRNA level for each condition was normalized to Gapdh mRNA level. F , Top, Experimental procedure. HEK293T cells expressing the nontarget shRNA or Baf53b shRNA were infected with either HSV-BAF53a or HSV-BAF53b ( n =3 for each condition). Middle, Representative Western blot. Bottom, BAF53a and BAF53b expression histograms. The y -axis indicates the BAF53a or BAF53b protein level relative to nontarget shRNA control. The BAF53a or BAF53b protein level for each condition was normalized to the GAPDH level. n values indicate the number of trials. G , Behavioral procedure for auditory fear conditioning with strong shock (0.5 mA). H , Top, Behavioral procedure. Bottom, Tone-induced freezing during retention test. Long-term memory tested 24 h after training in mice injected with either AAV-nontarget shRNA (control, n = 9) or AAV- Baf53b shRNA ( Baf53b KD, n = 15). n values indicate the number of mice. I , Top, Behavioral procedure. Bottom, Tone-induced freezing during retention test. Short-term memory tested 1 h after training (control, n = 11; Baf53b KD, n = 12). n values indicate the number of mice. J–L , Open field test with control ( n = 11) or Baf53b KD ( n = 10) mice. n values indicate the number of mice. J , Representative activity pattern tracked by EthoVision XT (Noldus). K , L , Center-crossings ( K ) and total distance moved ( L ) during 20 min. M , Construct maps for the AAV vectors expressing nontarget shRNA as a control and both knockdown-resistant BAF53b (rBAF53b) and Baf53b shRNA. Red represents mutation sites of rBAF53b. N , BAF53b expression levels from LA of noninfected mice (naive, n = 3), LAs infected with AAV-nontarget shRNA (nontarget shRNA, n = 3), AAV- Baf53b shRNA ( Baf53b shRNA, n = 3), or AAV-rBAF53b- Baf53b shRNA (rBAF53b, n = 3) as measured by Western blot. Top, Representative Western blot. Bottom, BAF53b expression histograms. The y -axis indicates the BAF53b protein level relative to the naive control. The BAF53b protein level for each condition was normalized to the GAPDH protein level. n values indicate the number of trials. O , Top, Behavioral procedure. Bottom, Freezing level measured in mice injected with either the AAV-nontarget shRNA (control, n = 6) or the AAV-rBAF53b- Baf53b shRNA ( n = 6) during long-term fear memory test. n values indicate the number of mice. Data are mean ± SEM. n.s., Not significant ( p > 0.05). * p
    Figure Legend Snippet: BAF53b function is necessary in the LA for the consolidation of auditory fear memory during long-term memory formation. A , Top, Construct map for AAV-nontarget shRNA and AAV- Baf53b shRNA. Bottom, Representative AAV-infected neurons (red) in the LA. Nuclei (blue) were stained with DAPI. Scale bar, 200 μm. B , Western blot showing BAF53b protein levels in the LA of naive mice ( n = 3), nontarget shRNA-expressing mice ( n = 3), and Baf53b shRNA-expressing mice ( n = 3). Top, Representative Western blot. Bottom, BAF53b expression histograms. The y -axis indicates the BAF53b protein level for each condition relative to naive control. The BAF53b protein level for each condition was normalized to the GAPDH level. n values indicate the number of trials. C , Western blot showing protein levels of BAF53a (left, anti-BAF53a) or BAF53b (right, anti-BAF53b) in HEK293T cells infected with either HSV-BAF53a or HSV-BAF53b. No infection condition was included as a control. GAPDH was used as a loading control. D , Top, Western blot showing BAF53b protein levels in the LA of Baf53b +/− (HET, n = 3) and WT ( n = 3) mice. Bottom, BAF53b expression histograms. The y -axis indicates the BAF53b protein level relative to the WT control. The BAF53b protein level for each condition was normalized to the GAPDH protein level. n values indicate the number of trials. E , Baf53b mRNA levels from LAs expressing either the nontarget shRNA ( n = 3) or the Baf53b shRNA ( n = 2) as measured by qPCR. The y -axis indicates the Baf53b mRNA level relative to the nontarget shRNA control. The Baf53b mRNA level for each condition was normalized to Gapdh mRNA level. F , Top, Experimental procedure. HEK293T cells expressing the nontarget shRNA or Baf53b shRNA were infected with either HSV-BAF53a or HSV-BAF53b ( n =3 for each condition). Middle, Representative Western blot. Bottom, BAF53a and BAF53b expression histograms. The y -axis indicates the BAF53a or BAF53b protein level relative to nontarget shRNA control. The BAF53a or BAF53b protein level for each condition was normalized to the GAPDH level. n values indicate the number of trials. G , Behavioral procedure for auditory fear conditioning with strong shock (0.5 mA). H , Top, Behavioral procedure. Bottom, Tone-induced freezing during retention test. Long-term memory tested 24 h after training in mice injected with either AAV-nontarget shRNA (control, n = 9) or AAV- Baf53b shRNA ( Baf53b KD, n = 15). n values indicate the number of mice. I , Top, Behavioral procedure. Bottom, Tone-induced freezing during retention test. Short-term memory tested 1 h after training (control, n = 11; Baf53b KD, n = 12). n values indicate the number of mice. J–L , Open field test with control ( n = 11) or Baf53b KD ( n = 10) mice. n values indicate the number of mice. J , Representative activity pattern tracked by EthoVision XT (Noldus). K , L , Center-crossings ( K ) and total distance moved ( L ) during 20 min. M , Construct maps for the AAV vectors expressing nontarget shRNA as a control and both knockdown-resistant BAF53b (rBAF53b) and Baf53b shRNA. Red represents mutation sites of rBAF53b. N , BAF53b expression levels from LA of noninfected mice (naive, n = 3), LAs infected with AAV-nontarget shRNA (nontarget shRNA, n = 3), AAV- Baf53b shRNA ( Baf53b shRNA, n = 3), or AAV-rBAF53b- Baf53b shRNA (rBAF53b, n = 3) as measured by Western blot. Top, Representative Western blot. Bottom, BAF53b expression histograms. The y -axis indicates the BAF53b protein level relative to the naive control. The BAF53b protein level for each condition was normalized to the GAPDH protein level. n values indicate the number of trials. O , Top, Behavioral procedure. Bottom, Freezing level measured in mice injected with either the AAV-nontarget shRNA (control, n = 6) or the AAV-rBAF53b- Baf53b shRNA ( n = 6) during long-term fear memory test. n values indicate the number of mice. Data are mean ± SEM. n.s., Not significant ( p > 0.05). * p

    Techniques Used: Construct, shRNA, Infection, Staining, Western Blot, Mouse Assay, Expressing, Real-time Polymerase Chain Reaction, Injection, Activity Assay, Mutagenesis

    BAF53b is induced in the LA after fear conditioning. A , Baf53b mRNA levels measured at intervals after auditory fear conditioning with strong shock (0.5 mA) and normalized to Gapdh (home cage, n = 7; 1 h, n = 4; 3 h, n = 4; 6 h, n = 8; 24 h, n = 5). The y -axis indicates the Baf53b mRNA level relative to the home cage control. The Baf53b mRNA level for each condition was normalized to Gapdh mRNA level. B , The absolute amount of Gapdh mRNA quantified using standard curve measured at different time points after auditory fear conditioning with strong shock (0.5 mA) (home cage, n = 7; 1 h, n = 4; 3 h, n = 4; 6 h, n = 8; 24 h, n = 5). The y -axis indicates the Gapdh mRNA level relative to the home cage control. n values indicate the number of mice. Data are mean ± SEM. * p
    Figure Legend Snippet: BAF53b is induced in the LA after fear conditioning. A , Baf53b mRNA levels measured at intervals after auditory fear conditioning with strong shock (0.5 mA) and normalized to Gapdh (home cage, n = 7; 1 h, n = 4; 3 h, n = 4; 6 h, n = 8; 24 h, n = 5). The y -axis indicates the Baf53b mRNA level relative to the home cage control. The Baf53b mRNA level for each condition was normalized to Gapdh mRNA level. B , The absolute amount of Gapdh mRNA quantified using standard curve measured at different time points after auditory fear conditioning with strong shock (0.5 mA) (home cage, n = 7; 1 h, n = 4; 3 h, n = 4; 6 h, n = 8; 24 h, n = 5). The y -axis indicates the Gapdh mRNA level relative to the home cage control. n values indicate the number of mice. Data are mean ± SEM. * p

    Techniques Used: Mouse Assay

    5) Product Images from "MicroRNA-210 suppresses glucocorticoid receptor expression in response to hypoxia in fetal rat cardiomyocytes"

    Article Title: MicroRNA-210 suppresses glucocorticoid receptor expression in response to hypoxia in fetal rat cardiomyocytes

    Journal: Oncotarget

    doi: 10.18632/oncotarget.17801

    Hypoxia induces miR-210 expression and suppresses GR expression in fetal rat hearts ( A ) Hearts were isolated from E21 fetuses from pregnant rats treated with normoxia or hypoxia at 10.5% O 2 from day 15 to day 21 of gestation, and miR-210 expression was measured by miScript miR real-time RT-qPCR. n = 8. ( B , C , D ) Cardiomyocytes isolated from E21 fetuses were treated with normoxia or hypoxia (1% O 2 ) for 24 hours. MiR-210 expression was measured by miScript miR real-time qRT-PCR (B), n = 6; GR protein abundance was measured by Western blot (C), n = 4–5, and GR mRNA abundance was determined by real-time qRT-PCR (D), n = 6. Data are mean ± SEM. * p
    Figure Legend Snippet: Hypoxia induces miR-210 expression and suppresses GR expression in fetal rat hearts ( A ) Hearts were isolated from E21 fetuses from pregnant rats treated with normoxia or hypoxia at 10.5% O 2 from day 15 to day 21 of gestation, and miR-210 expression was measured by miScript miR real-time RT-qPCR. n = 8. ( B , C , D ) Cardiomyocytes isolated from E21 fetuses were treated with normoxia or hypoxia (1% O 2 ) for 24 hours. MiR-210 expression was measured by miScript miR real-time qRT-PCR (B), n = 6; GR protein abundance was measured by Western blot (C), n = 4–5, and GR mRNA abundance was determined by real-time qRT-PCR (D), n = 6. Data are mean ± SEM. * p

    Techniques Used: Expressing, Isolation, Quantitative RT-PCR, Western Blot

    6) Product Images from "MicroRNA Profiling Identifies miR-196a as Differentially Expressed in Childhood Adrenoleukodystrophy and Adult Adrenomyeloneuropathy"

    Article Title: MicroRNA Profiling Identifies miR-196a as Differentially Expressed in Childhood Adrenoleukodystrophy and Adult Adrenomyeloneuropathy

    Journal: Molecular neurobiology

    doi: 10.1007/s12035-016-9746-0

    Validation of miR-196a expression in human skin fibroblasts ( a ) and human brain samples ( b ) by qRT-PCR. miR-196a expression was validated in human skin fibroblasts and human brain samples (from equivalent brain regions) using Qiagen miScript primer assays by qRT-PCR. The values were normalized to SNORD61 and presented as fold change. As represented by figure, miR-196a expression was downregulated in cALD fibroblasts and brain samples and upregulated in AMN fibroblasts and brain samples. B3 is brain sample from AMN that subsequently developed CNS disease. Downregulation of miR-196a expression in brain sample B3 indicates of cerebral involvement. Thirty-five percent of AMN patients develop cerebral involvement
    Figure Legend Snippet: Validation of miR-196a expression in human skin fibroblasts ( a ) and human brain samples ( b ) by qRT-PCR. miR-196a expression was validated in human skin fibroblasts and human brain samples (from equivalent brain regions) using Qiagen miScript primer assays by qRT-PCR. The values were normalized to SNORD61 and presented as fold change. As represented by figure, miR-196a expression was downregulated in cALD fibroblasts and brain samples and upregulated in AMN fibroblasts and brain samples. B3 is brain sample from AMN that subsequently developed CNS disease. Downregulation of miR-196a expression in brain sample B3 indicates of cerebral involvement. Thirty-five percent of AMN patients develop cerebral involvement

    Techniques Used: Expressing, Quantitative RT-PCR

    Roles of miR-196a in target gene expression. To establish the regulation of target genes by miR-196a, miR-196a mimic was transfected in ABCD1 knockdown U87 astrocytes cells (U87 Lenti) and transfection efficiency evaluated by miScript qRT-PCR. SNORD61 was used as an internal control and represented as miR-196a/SNORD61 ( a ). The expression of target genes IKKα and IKKβ was analysed after the transfection of miR-196a mimic and normalized to GAPDH. The expression of these genes was significantly increased in ABCD1 silenced cells as compared to wild type but significantly decreased after transfection of mimic in U87 Lenti cells ( b and c ). ** P
    Figure Legend Snippet: Roles of miR-196a in target gene expression. To establish the regulation of target genes by miR-196a, miR-196a mimic was transfected in ABCD1 knockdown U87 astrocytes cells (U87 Lenti) and transfection efficiency evaluated by miScript qRT-PCR. SNORD61 was used as an internal control and represented as miR-196a/SNORD61 ( a ). The expression of target genes IKKα and IKKβ was analysed after the transfection of miR-196a mimic and normalized to GAPDH. The expression of these genes was significantly increased in ABCD1 silenced cells as compared to wild type but significantly decreased after transfection of mimic in U87 Lenti cells ( b and c ). ** P

    Techniques Used: Expressing, Transfection, Quantitative RT-PCR

    7) Product Images from "The effects of microRNAs on human neural stem cell differentiation in two- and three-dimensional cultures"

    Article Title: The effects of microRNAs on human neural stem cell differentiation in two- and three-dimensional cultures

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/scrt437

    Human cell differentiation and development miScript miRNA PCR array profiles. Group clustergram analysis of miRNA PCR array profiles analysis of differentially regulated miRNAs identified in control (undifferentiated hNSCs) and hNSCs seeded on 2D and 3D substrates and differentiated for 1W and 3W.
    Figure Legend Snippet: Human cell differentiation and development miScript miRNA PCR array profiles. Group clustergram analysis of miRNA PCR array profiles analysis of differentially regulated miRNAs identified in control (undifferentiated hNSCs) and hNSCs seeded on 2D and 3D substrates and differentiated for 1W and 3W.

    Techniques Used: Cell Differentiation, Polymerase Chain Reaction

    8) Product Images from "miR-34c-5p and CaMKII are involved in aldosterone-induced fibrosis in kidney collecting duct cells"

    Article Title: miR-34c-5p and CaMKII are involved in aldosterone-induced fibrosis in kidney collecting duct cells

    Journal: American Journal of Physiology - Renal Physiology

    doi: 10.1152/ajprenal.00358.2017

    Changes of CaMKIIβ and fibronectin (FN) protein expression in mpkCCDc14 cells. A and B : semiquantitative immunoblotting of CaMKIIβ (~58 and 62 kDa; arrows) in mpkCCDc14 cells with CaMKIIβ siRNA-mediated knockdown in the absence of aldosterone treatment. C and D : semiquantitative immunoblotting of CaMKIIβ (~58 and 62 kDa; arrows) and FN (~262 kDa) in mpkCCDc14 cells with siRNA-mediated knockdown of CaMKIIβ (40 nM), followed by aldosterone treatment (10 −6 M; 3 days). E and F : semiquantitative immunoblotting of CaMKIIβ (~58 and 62 kDa; arrows) and FN (~262 kDa) in mpkCCDc14 cells transfected with miR-34c-5p mimic (20 nM), followed by aldosterone treatment (10 −6 M; 3 days). G : real-time quantitative PCR analysis for the change of CaMKIIβ mRNA expression in mpkCCDc14 cells transfected with the miR-34c-5p mimic (20 nM), followed by aldosterone treatment (10 −6 M; 3 days). Aldo, aldosterone; Con siRNA, nontarget control siRNA; NC, miScript Inhibitor Negative Control; n, number of cell lysate preparation. * P
    Figure Legend Snippet: Changes of CaMKIIβ and fibronectin (FN) protein expression in mpkCCDc14 cells. A and B : semiquantitative immunoblotting of CaMKIIβ (~58 and 62 kDa; arrows) in mpkCCDc14 cells with CaMKIIβ siRNA-mediated knockdown in the absence of aldosterone treatment. C and D : semiquantitative immunoblotting of CaMKIIβ (~58 and 62 kDa; arrows) and FN (~262 kDa) in mpkCCDc14 cells with siRNA-mediated knockdown of CaMKIIβ (40 nM), followed by aldosterone treatment (10 −6 M; 3 days). E and F : semiquantitative immunoblotting of CaMKIIβ (~58 and 62 kDa; arrows) and FN (~262 kDa) in mpkCCDc14 cells transfected with miR-34c-5p mimic (20 nM), followed by aldosterone treatment (10 −6 M; 3 days). G : real-time quantitative PCR analysis for the change of CaMKIIβ mRNA expression in mpkCCDc14 cells transfected with the miR-34c-5p mimic (20 nM), followed by aldosterone treatment (10 −6 M; 3 days). Aldo, aldosterone; Con siRNA, nontarget control siRNA; NC, miScript Inhibitor Negative Control; n, number of cell lysate preparation. * P

    Techniques Used: Expressing, Transfection, Real-time Polymerase Chain Reaction, Negative Control

    9) Product Images from "Original Research: Stable expression of miR-34a mediates fetal hemoglobin induction in K562 cells"

    Article Title: Original Research: Stable expression of miR-34a mediates fetal hemoglobin induction in K562 cells

    Journal: Experimental Biology and Medicine

    doi: 10.1177/1535370216636725

    miR-34a overexpression mediates HbF induction in K562 stable pools. K562 cells were transduced with the SMARTchoice™ shMIMIC miR-34a or scrambled lentivirus particles. After puromycin selection, cells were harvested at day 9 and day 16 for analysis (see Materials and methods) (a) Mature miR-34a expression was measured by RT-qPCR using the Qiagen’s miScript Primer Assay System. The ratio of miR-34a to RNU6 (housekeeping control) was plotted. The data were normalized to scramble control and reported as fold change of the mean ± standard error of the mean; * p
    Figure Legend Snippet: miR-34a overexpression mediates HbF induction in K562 stable pools. K562 cells were transduced with the SMARTchoice™ shMIMIC miR-34a or scrambled lentivirus particles. After puromycin selection, cells were harvested at day 9 and day 16 for analysis (see Materials and methods) (a) Mature miR-34a expression was measured by RT-qPCR using the Qiagen’s miScript Primer Assay System. The ratio of miR-34a to RNU6 (housekeeping control) was plotted. The data were normalized to scramble control and reported as fold change of the mean ± standard error of the mean; * p

    Techniques Used: Over Expression, Transduction, Selection, Expressing, Quantitative RT-PCR

    10) Product Images from "miR-214 promotes periodontal ligament stem cell osteoblastic differentiation by modulating Wnt/β-catenin signaling"

    Article Title: miR-214 promotes periodontal ligament stem cell osteoblastic differentiation by modulating Wnt/β-catenin signaling

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2017.7821

    miRNA expression in differentiated and non-differentiated PDLSCs. (A) Heat map of miRNA profiles represents the differentially expressed miRNAs between differentiated and non-differentiated PDLSCs. Green indicates low expression levels; red indicates high expression levels. (B) RT-qPCR was performed to determine the expression of miRNAs in differentiated and undifferentiated PDLSCs. (C) RT-qPCR was used to assess the expression levels of miR-214 at the indicated time points in differentiated and non-differentiated PDLSCs. miR-214 expression was normalized to U6 small nuclear RNA. **P
    Figure Legend Snippet: miRNA expression in differentiated and non-differentiated PDLSCs. (A) Heat map of miRNA profiles represents the differentially expressed miRNAs between differentiated and non-differentiated PDLSCs. Green indicates low expression levels; red indicates high expression levels. (B) RT-qPCR was performed to determine the expression of miRNAs in differentiated and undifferentiated PDLSCs. (C) RT-qPCR was used to assess the expression levels of miR-214 at the indicated time points in differentiated and non-differentiated PDLSCs. miR-214 expression was normalized to U6 small nuclear RNA. **P

    Techniques Used: Expressing, Quantitative RT-PCR

    11) Product Images from "miR-10a Regulates Proliferation of Human Cardiomyocyte Progenitor Cells by Targeting GATA6"

    Article Title: miR-10a Regulates Proliferation of Human Cardiomyocyte Progenitor Cells by Targeting GATA6

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0103097

    miR-10a does not influence hCMPC differentiation toward cardiomyocytes. A. Representative image of differentiating hCMPCs with manipulated expression of miR-10a or mock. Cells were stained with DAPI, α-actinin and Trop I (×200). B. Data collected from A. *P
    Figure Legend Snippet: miR-10a does not influence hCMPC differentiation toward cardiomyocytes. A. Representative image of differentiating hCMPCs with manipulated expression of miR-10a or mock. Cells were stained with DAPI, α-actinin and Trop I (×200). B. Data collected from A. *P

    Techniques Used: Expressing, Staining

    Expression of miR-10a is increased in cardiogenesis. Hearts of different stages of cardiogenesis were collected for miR-10a expression analysis. Quantification of miR-10a level was related to U6. *P
    Figure Legend Snippet: Expression of miR-10a is increased in cardiogenesis. Hearts of different stages of cardiogenesis were collected for miR-10a expression analysis. Quantification of miR-10a level was related to U6. *P

    Techniques Used: Expressing

    miR-10a binds to GATA6 and suppresses GATA6 expression. A. Pmir- glo dual luciferase plasmid containing a wild type or mutant GATA6-3′UTR was cotransfected with miR-10a mimics or mock into 293 cells. The relative luciferase activity was shown. *P
    Figure Legend Snippet: miR-10a binds to GATA6 and suppresses GATA6 expression. A. Pmir- glo dual luciferase plasmid containing a wild type or mutant GATA6-3′UTR was cotransfected with miR-10a mimics or mock into 293 cells. The relative luciferase activity was shown. *P

    Techniques Used: Expressing, Luciferase, Plasmid Preparation, Mutagenesis, Activity Assay

    miR-10a reduces proliferation of hCMPCs. A. miR-10a mimics decrease the BrdU incorporation rate of hCMPCs, whereas a miR-10a inhibitor does not significantly affect the process. *P
    Figure Legend Snippet: miR-10a reduces proliferation of hCMPCs. A. miR-10a mimics decrease the BrdU incorporation rate of hCMPCs, whereas a miR-10a inhibitor does not significantly affect the process. *P

    Techniques Used: BrdU Incorporation Assay

    12) Product Images from "Analysis of circulating-microRNA expression in lactating Holstein cows under summer heat stress"

    Article Title: Analysis of circulating-microRNA expression in lactating Holstein cows under summer heat stress

    Journal: bioRxiv

    doi: 10.1101/2020.03.18.996777

    Volcano plot showing differentially expressed miRNAs between NHS and HS using transformed normalized data. |Fold change| value ≥ 2 and P
    Figure Legend Snippet: Volcano plot showing differentially expressed miRNAs between NHS and HS using transformed normalized data. |Fold change| value ≥ 2 and P

    Techniques Used: Transformation Assay

    13) Product Images from "Over-expression of transcription factor ARK1 gene leads to down-regulation of lignin synthesis related genes in hybrid poplar ‘717’"

    Article Title: Over-expression of transcription factor ARK1 gene leads to down-regulation of lignin synthesis related genes in hybrid poplar ‘717’

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-65328-y

    Comparison of RNA-sequencing and RT-qPCR results of selected DEGs. , RNA-seq; ◼, RT-qPCR. 1, Cyclin-D3-1 protein; 2, PYR1 abscisic acid receptor; 3, Histidine phosphate transfer protein; 4, Indole-3-acetic amido synthetase; 5, Indole-3-acetic amido synthase GH3.6; 6, indole-3-acetic amido synthase GH3.9; 7, Trans cinnamic acid-hydroxylase; 8, Peroxidase 17; 9, Peroxidase 73. RT-qPCR was performed on 3 transgenic lines used in the transcriptome analysis and 3 control seedlings, normalized with housekeeping gene EF1β, repeated 3 times. DEG, differentially expressed gene; RT-qPCR, Real time quantitative polymerase chain reaction.
    Figure Legend Snippet: Comparison of RNA-sequencing and RT-qPCR results of selected DEGs. , RNA-seq; ◼, RT-qPCR. 1, Cyclin-D3-1 protein; 2, PYR1 abscisic acid receptor; 3, Histidine phosphate transfer protein; 4, Indole-3-acetic amido synthetase; 5, Indole-3-acetic amido synthase GH3.6; 6, indole-3-acetic amido synthase GH3.9; 7, Trans cinnamic acid-hydroxylase; 8, Peroxidase 17; 9, Peroxidase 73. RT-qPCR was performed on 3 transgenic lines used in the transcriptome analysis and 3 control seedlings, normalized with housekeeping gene EF1β, repeated 3 times. DEG, differentially expressed gene; RT-qPCR, Real time quantitative polymerase chain reaction.

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

    14) Product Images from "Over-expression of transcription factor ARK1 gene leads to down-regulation of lignin synthesis related genes in hybrid poplar ‘717’"

    Article Title: Over-expression of transcription factor ARK1 gene leads to down-regulation of lignin synthesis related genes in hybrid poplar ‘717’

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-65328-y

    Comparison of RNA-sequencing and RT-qPCR results of selected DEGs. , RNA-seq; ◼, RT-qPCR. 1, Cyclin-D3-1 protein; 2, PYR1 abscisic acid receptor; 3, Histidine phosphate transfer protein; 4, Indole-3-acetic amido synthetase; 5, Indole-3-acetic amido synthase GH3.6; 6, indole-3-acetic amido synthase GH3.9; 7, Trans cinnamic acid-hydroxylase; 8, Peroxidase 17; 9, Peroxidase 73. RT-qPCR was performed on 3 transgenic lines used in the transcriptome analysis and 3 control seedlings, normalized with housekeeping gene EF1β, repeated 3 times. DEG, differentially expressed gene; RT-qPCR, Real time quantitative polymerase chain reaction.
    Figure Legend Snippet: Comparison of RNA-sequencing and RT-qPCR results of selected DEGs. , RNA-seq; ◼, RT-qPCR. 1, Cyclin-D3-1 protein; 2, PYR1 abscisic acid receptor; 3, Histidine phosphate transfer protein; 4, Indole-3-acetic amido synthetase; 5, Indole-3-acetic amido synthase GH3.6; 6, indole-3-acetic amido synthase GH3.9; 7, Trans cinnamic acid-hydroxylase; 8, Peroxidase 17; 9, Peroxidase 73. RT-qPCR was performed on 3 transgenic lines used in the transcriptome analysis and 3 control seedlings, normalized with housekeeping gene EF1β, repeated 3 times. DEG, differentially expressed gene; RT-qPCR, Real time quantitative polymerase chain reaction.

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

    15) Product Images from "Epigenetic alterations in hippocampus of SAMP8 senescent mice and modulation by voluntary physical exercise"

    Article Title: Epigenetic alterations in hippocampus of SAMP8 senescent mice and modulation by voluntary physical exercise

    Journal: Frontiers in Aging Neuroscience

    doi: 10.3389/fnagi.2014.00051

    Hippocampal microRNAs modulated by exercise in SAMR1 and SAMP8 mice . MicroRNA expression was measured using miScript ® miRNA PCR Array- Neurological Development and Disease miRNA PCR Array (Qiagen) and expressed relative to housekeeping miRNAs proposed by the array. (A–C) miRNAs that are altered in sedentary SAMP8 compared with sedentary SAMR1 mice and regulated by exercise in senescent mice, (miR-28a-5p, miR-98-5p, and mir-148b-3p, respectively). (D–G) miRNAs unaltered in sedentary SAMP8 mice but responsive to exercise intervention in SAMR1 and SAMP8 mice (miR-7a-5p, miR-15b-5p, miR-105, and miR-133b-3p, respectively). Means ± standard error are represented; Two-Way ANOVA results are indicated as * p
    Figure Legend Snippet: Hippocampal microRNAs modulated by exercise in SAMR1 and SAMP8 mice . MicroRNA expression was measured using miScript ® miRNA PCR Array- Neurological Development and Disease miRNA PCR Array (Qiagen) and expressed relative to housekeeping miRNAs proposed by the array. (A–C) miRNAs that are altered in sedentary SAMP8 compared with sedentary SAMR1 mice and regulated by exercise in senescent mice, (miR-28a-5p, miR-98-5p, and mir-148b-3p, respectively). (D–G) miRNAs unaltered in sedentary SAMP8 mice but responsive to exercise intervention in SAMR1 and SAMP8 mice (miR-7a-5p, miR-15b-5p, miR-105, and miR-133b-3p, respectively). Means ± standard error are represented; Two-Way ANOVA results are indicated as * p

    Techniques Used: Mouse Assay, Expressing, Polymerase Chain Reaction

    16) Product Images from "Gastrin-induced miR-222 promotes gastric tumor development by suppressing p27kip1"

    Article Title: Gastrin-induced miR-222 promotes gastric tumor development by suppressing p27kip1

    Journal: Oncotarget

    doi: 10.18632/oncotarget.9990

    In AGS GR cells treated with 10nM G17 compared with untreated controls, miScript miRNA PCR arrays showed 3 miRNAs that increased and 3 miRNAs that decreased in expression beyond the 2-fold threshold, with only miR-222 and miR-376c proving significant ( A ) As the abundance of miR-376c was low in both the treated and untreated samples, miR-222 was further investigated ( B ). miR-222 expression increased dose and time dependently in AGS GR cells and was maximal following treatment with 10nM G17 for 24h in serum free media. miR-222 expression did not significantly change following G17 treatment of untransfected AGS cells ( C , D ). LY294002 (20 μM), YM022 (100 nM), netazepide (100 nM) and Ro-32-0432 (1 μM) all completely reversed while PD98089 (20 μM) partially reversed the miR-222 overexpression caused by 10 nM G17 treatment of AGS GR cells for 24 h ( E ). Ro-32-0432 (1 μM) also completely reversed while LY294002 (20 μM) partially reversed the miR-222 overexpression induced by 100 nM PMA treatment of the same cell line for 24 h ( F ). Statistical significance was determined using two-way ANOVA with Sidak post-hoc test and P
    Figure Legend Snippet: In AGS GR cells treated with 10nM G17 compared with untreated controls, miScript miRNA PCR arrays showed 3 miRNAs that increased and 3 miRNAs that decreased in expression beyond the 2-fold threshold, with only miR-222 and miR-376c proving significant ( A ) As the abundance of miR-376c was low in both the treated and untreated samples, miR-222 was further investigated ( B ). miR-222 expression increased dose and time dependently in AGS GR cells and was maximal following treatment with 10nM G17 for 24h in serum free media. miR-222 expression did not significantly change following G17 treatment of untransfected AGS cells ( C , D ). LY294002 (20 μM), YM022 (100 nM), netazepide (100 nM) and Ro-32-0432 (1 μM) all completely reversed while PD98089 (20 μM) partially reversed the miR-222 overexpression caused by 10 nM G17 treatment of AGS GR cells for 24 h ( E ). Ro-32-0432 (1 μM) also completely reversed while LY294002 (20 μM) partially reversed the miR-222 overexpression induced by 100 nM PMA treatment of the same cell line for 24 h ( F ). Statistical significance was determined using two-way ANOVA with Sidak post-hoc test and P

    Techniques Used: Polymerase Chain Reaction, Expressing, Over Expression

    17) Product Images from "FTY720/Fingolimod Reduces Synucleinopathy and Improves Gut Motility in A53T Mice"

    Article Title: FTY720/Fingolimod Reduces Synucleinopathy and Improves Gut Motility in A53T Mice

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M116.744029

    FTY720 stimulates long term increases in BDNF protein in aging Tg mice in association with significantly lower levels of miR206–3p. A , BDNF protein normalized to β-actin on immunoblots confirmed that BDNF was increased in colons of FTY720-treated 21-month-old mice. B , the expression of the regulatory microRNA, miR206–3p, was significantly lower in response to FTY720 treatment of aged Tg mice as compared with vehicle Tg mice. The decrease in miR206–3p was further validated in a control experiment with dopaminergic MN9D cells treated with 160 n m FTY720 for 24 h. ( n = 8 mice/treatment group); *, p
    Figure Legend Snippet: FTY720 stimulates long term increases in BDNF protein in aging Tg mice in association with significantly lower levels of miR206–3p. A , BDNF protein normalized to β-actin on immunoblots confirmed that BDNF was increased in colons of FTY720-treated 21-month-old mice. B , the expression of the regulatory microRNA, miR206–3p, was significantly lower in response to FTY720 treatment of aged Tg mice as compared with vehicle Tg mice. The decrease in miR206–3p was further validated in a control experiment with dopaminergic MN9D cells treated with 160 n m FTY720 for 24 h. ( n = 8 mice/treatment group); *, p

    Techniques Used: Mouse Assay, Western Blot, Expressing

    18) Product Images from "Na/K-ATPase signaling mediates miR-29b-3p regulation and cardiac fibrosis formation in mice with chronic kidney disease"

    Article Title: Na/K-ATPase signaling mediates miR-29b-3p regulation and cardiac fibrosis formation in mice with chronic kidney disease

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0197688

    The effect of NFκB inhibitor on ouabain-induced miR-29b-3p regulation in cardiac fibroblasts isolated from WT and α1+/- mice. Isolated primary cultures of mouse cardiac fibroblasts were pretreated with 1μM of the NFκB inhibitor BAY 11–7082 for 15 min followed by ouabain (10 or 100nM) treatment for 24h. Non-treated or ouabain treatment alone without BAY 11–7082 was used as control. After treatment, cell lysates were collected. Expression of miR-29b-3p was measured using RT-qPCR in RNA isolated from cells from WT animals ( A ) and α1 +/- animals ( B ). Fibroblasts were were obtained from 4 animals in each group. Data was analyzed using One-Way ANOVA with GraphPad software 7.0. * indicates p
    Figure Legend Snippet: The effect of NFκB inhibitor on ouabain-induced miR-29b-3p regulation in cardiac fibroblasts isolated from WT and α1+/- mice. Isolated primary cultures of mouse cardiac fibroblasts were pretreated with 1μM of the NFκB inhibitor BAY 11–7082 for 15 min followed by ouabain (10 or 100nM) treatment for 24h. Non-treated or ouabain treatment alone without BAY 11–7082 was used as control. After treatment, cell lysates were collected. Expression of miR-29b-3p was measured using RT-qPCR in RNA isolated from cells from WT animals ( A ) and α1 +/- animals ( B ). Fibroblasts were were obtained from 4 animals in each group. Data was analyzed using One-Way ANOVA with GraphPad software 7.0. * indicates p

    Techniques Used: Isolation, Mouse Assay, Expressing, Quantitative RT-PCR, Software

    PNx-induced miR-29b-3p expression change in WT and α1+/- mice. WT and α1+/- mice were subjected to sham or 5/6 th partial nephrectomy (PNx) surgery and left ventricle tissue was collected at the end of 16 th week for RNA extraction, RT-qPCR, and Western blot analyses. A): Western blot of Na/K-ATPase α1 subunit expression in left ventricle tissue from different group of mice. !! indicates p
    Figure Legend Snippet: PNx-induced miR-29b-3p expression change in WT and α1+/- mice. WT and α1+/- mice were subjected to sham or 5/6 th partial nephrectomy (PNx) surgery and left ventricle tissue was collected at the end of 16 th week for RNA extraction, RT-qPCR, and Western blot analyses. A): Western blot of Na/K-ATPase α1 subunit expression in left ventricle tissue from different group of mice. !! indicates p

    Techniques Used: Expressing, Mouse Assay, RNA Extraction, Quantitative RT-PCR, Western Blot

    Injection of pNaKtide diminishes PNx-induced decrease in miR-29b-3p expression. Total RNA obtained from left ventricle tissue were used for RT-qPCR analyses. Expression of miR-29b-3p was presented as fold regulation relevant to WT sham animals. Data were obtained from 5 animals in each group and was analyzed using Two-Way ANOVA with GraphPad software 7.0. * indicates p
    Figure Legend Snippet: Injection of pNaKtide diminishes PNx-induced decrease in miR-29b-3p expression. Total RNA obtained from left ventricle tissue were used for RT-qPCR analyses. Expression of miR-29b-3p was presented as fold regulation relevant to WT sham animals. Data were obtained from 5 animals in each group and was analyzed using Two-Way ANOVA with GraphPad software 7.0. * indicates p

    Techniques Used: Injection, Expressing, Quantitative RT-PCR, Software

    19) Product Images from "MicroRNA-141-3p Negatively Modulates SDF-1 Expression in Age-Dependent Pathophysiology of Human and Murine Bone Marrow Stromal Cells"

    Article Title: MicroRNA-141-3p Negatively Modulates SDF-1 Expression in Age-Dependent Pathophysiology of Human and Murine Bone Marrow Stromal Cells

    Journal: The Journals of Gerontology Series A: Biological Sciences and Medical Sciences

    doi: 10.1093/gerona/gly186

    Luciferase reporter assays demonstrating the SDF1 3′-UTR is the target of miR-141-3p. ( a ) Schematic representation of the 3′-UTR of human SDF1 mRNA with the two putative complementary sequences to miR-141 and interspecies conservation of putative miR-141 binding sites within the SDF1 ( b ) Luciferase activity assays detecting the miR-141-3p binding sites on the 3′-UTR of SDF1. Cotransfection of control miRNA/siRNA and luciferase reporters with the wild-type SDF1 3′-UTR was used as a control. In other groups, miR-141-3p mimics were cotransfected with luciferase reporters containing wild-type 3′-UTR or the 3′-UTR with mutated binding site 1 (M1/positions 51–58), with mutated binding site 2 (M2/positions 2940–2947), and with both mutations (M3). The control mimic was independently set to 100%. Data are presented as mean ± SEM ( n = 5; # p
    Figure Legend Snippet: Luciferase reporter assays demonstrating the SDF1 3′-UTR is the target of miR-141-3p. ( a ) Schematic representation of the 3′-UTR of human SDF1 mRNA with the two putative complementary sequences to miR-141 and interspecies conservation of putative miR-141 binding sites within the SDF1 ( b ) Luciferase activity assays detecting the miR-141-3p binding sites on the 3′-UTR of SDF1. Cotransfection of control miRNA/siRNA and luciferase reporters with the wild-type SDF1 3′-UTR was used as a control. In other groups, miR-141-3p mimics were cotransfected with luciferase reporters containing wild-type 3′-UTR or the 3′-UTR with mutated binding site 1 (M1/positions 51–58), with mutated binding site 2 (M2/positions 2940–2947), and with both mutations (M3). The control mimic was independently set to 100%. Data are presented as mean ± SEM ( n = 5; # p

    Techniques Used: Luciferase, Binding Assay, Activity Assay, Cotransfection

    miR-141-3p reduces SDF1 in mice and human BMSCs: ( a ) Mouse and ( b ) human BMSCs were transfected with miRNA mimic miR-141 and NC control (nontargeting siRNAs) for 48 hours. These cells were then used for RNA isolation. Data (means ± SD , n = 5) are represented as the fold change in expression compared with control (* p
    Figure Legend Snippet: miR-141-3p reduces SDF1 in mice and human BMSCs: ( a ) Mouse and ( b ) human BMSCs were transfected with miRNA mimic miR-141 and NC control (nontargeting siRNAs) for 48 hours. These cells were then used for RNA isolation. Data (means ± SD , n = 5) are represented as the fold change in expression compared with control (* p

    Techniques Used: Mouse Assay, Transfection, Isolation, Expressing

    Age-related dysregulation of miR-141-3p on mouse and human bone marrow niche. Real-time PCR of miRNA-141-3p on ( a ) young ( n = 6) and old ( n = 5) mouse bone marrow interstitial fluid and ( b ) young age 18–40 ( n = 9) and old age 60 and above ( n = 11) human bone marrow stromal cells. Data (means ± SD ) are represented as the fold change in expression compared with young (* p value = .05).
    Figure Legend Snippet: Age-related dysregulation of miR-141-3p on mouse and human bone marrow niche. Real-time PCR of miRNA-141-3p on ( a ) young ( n = 6) and old ( n = 5) mouse bone marrow interstitial fluid and ( b ) young age 18–40 ( n = 9) and old age 60 and above ( n = 11) human bone marrow stromal cells. Data (means ± SD ) are represented as the fold change in expression compared with young (* p value = .05).

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing

    20) Product Images from "Novel MIR143-NOTCH Fusions in Benign and Malignant Glomus Tumors"

    Article Title: Novel MIR143-NOTCH Fusions in Benign and Malignant Glomus Tumors

    Journal: Genes, chromosomes & cancer

    doi: 10.1002/gcc.22102

    MIR143-NOTCH2 gene fusion in a malignant gastrointestinal glomus tumor (GT1)
    Figure Legend Snippet: MIR143-NOTCH2 gene fusion in a malignant gastrointestinal glomus tumor (GT1)

    Techniques Used:

    MIR143-NOTCH1 gene fusion in a benign glomus tumor of the neck soft tissue (GT2)
    Figure Legend Snippet: MIR143-NOTCH1 gene fusion in a benign glomus tumor of the neck soft tissue (GT2)

    Techniques Used:

    MIR143-NOTCH3 fusion in a benign glomus tumor of the forearm (GT19)
    Figure Legend Snippet: MIR143-NOTCH3 fusion in a benign glomus tumor of the forearm (GT19)

    Techniques Used:

    MIR143-NOTCH2 fusion results in overexpression of 3′- NOTCH2 mRNA, triggered by the a strong MIR143 promoter, which is highly expressed in smooth muscle lineage
    Figure Legend Snippet: MIR143-NOTCH2 fusion results in overexpression of 3′- NOTCH2 mRNA, triggered by the a strong MIR143 promoter, which is highly expressed in smooth muscle lineage

    Techniques Used: Over Expression

    21) Product Images from "An Expanded Role for HLA Genes: HLA-B Encodes a microRNA that Regulates IgA and Other Immune Response Transcripts"

    Article Title: An Expanded Role for HLA Genes: HLA-B Encodes a microRNA that Regulates IgA and Other Immune Response Transcripts

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2017.00583

    HLA-B intron 4 sequence variability and miR-6891 isomiR characterization . (A) There are 384 annotated HLA-B alleles with full-length sequence annotation within the ImMunoGeneTics (IMGT) database (release 3.25), with each allele represented by one of eight unique intron 4 sequence motifs. The aligned sequence motifs are provided along with their allele frequency within IMGT and polymorphic positions (highlighted in red). (B) Sequence logo plot depicting the lack of polymorphism within HSA-miR-6891-5p. (C) Sequence logo plot depicting polymorphic sites within HSA-miR-6891-3p at positions 6 and 14 of the mature miRNA.
    Figure Legend Snippet: HLA-B intron 4 sequence variability and miR-6891 isomiR characterization . (A) There are 384 annotated HLA-B alleles with full-length sequence annotation within the ImMunoGeneTics (IMGT) database (release 3.25), with each allele represented by one of eight unique intron 4 sequence motifs. The aligned sequence motifs are provided along with their allele frequency within IMGT and polymorphic positions (highlighted in red). (B) Sequence logo plot depicting the lack of polymorphism within HSA-miR-6891-5p. (C) Sequence logo plot depicting polymorphic sites within HSA-miR-6891-3p at positions 6 and 14 of the mature miRNA.

    Techniques Used: Sequencing

    Exploring the role of miR-6891-5p in selective IgA deficiency (A) pedigree of affected (proband, black shadowing) and unaffected (white shadowing) family members is presented in panels B and C . (B) HLA-B , miR-6891-5p IGHA1 , and IGHA2 expression (qPCR) among IgA-deficient B-LCLs collected from affected individuals and unaffected family members (standard error bars shown, n = 3). (C) Selective IgA deficient cell line ID18 was transduced with a lentiviral construct expressing either the antisense miR-6891-5p (miR-6891-5p inhibition) or the scrambled sequence of antisense miR-6891-5p (control). Total RNA was purified, and IGHA1 and IGHA2 mRNA transcript levels were analyzed by qPCR ( y -axis shown on left of plot, standard error bars shown, n = 3). After 24 h, media was collected and analyzed by ELISA using anti-IgA antibody ( y -axis shown on right of plot, standard error bars shown, n = 3). All p -values shown are calculated using a t -test.
    Figure Legend Snippet: Exploring the role of miR-6891-5p in selective IgA deficiency (A) pedigree of affected (proband, black shadowing) and unaffected (white shadowing) family members is presented in panels B and C . (B) HLA-B , miR-6891-5p IGHA1 , and IGHA2 expression (qPCR) among IgA-deficient B-LCLs collected from affected individuals and unaffected family members (standard error bars shown, n = 3). (C) Selective IgA deficient cell line ID18 was transduced with a lentiviral construct expressing either the antisense miR-6891-5p (miR-6891-5p inhibition) or the scrambled sequence of antisense miR-6891-5p (control). Total RNA was purified, and IGHA1 and IGHA2 mRNA transcript levels were analyzed by qPCR ( y -axis shown on left of plot, standard error bars shown, n = 3). After 24 h, media was collected and analyzed by ELISA using anti-IgA antibody ( y -axis shown on right of plot, standard error bars shown, n = 3). All p -values shown are calculated using a t -test.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Transduction, Construct, Inhibition, Sequencing, Purification, Enzyme-linked Immunosorbent Assay

    Predicted biogenesis of HSA-miR-6891 . miR-6891 is derived from intron 4 of HLA-B , which upon exon splicing of the HLA-B transcript forms a stable pre-miRNA hairpin structure. The pre-miRNA is then processed by the Dicer enzyme to form two mature miRNA products, HSA-miR-6891-5p and HSA-miR-6891-3p.
    Figure Legend Snippet: Predicted biogenesis of HSA-miR-6891 . miR-6891 is derived from intron 4 of HLA-B , which upon exon splicing of the HLA-B transcript forms a stable pre-miRNA hairpin structure. The pre-miRNA is then processed by the Dicer enzyme to form two mature miRNA products, HSA-miR-6891-5p and HSA-miR-6891-3p.

    Techniques Used: Derivative Assay

    Identification of potential miR-6891-5p targets . COX cells were transduced with lentiviruses expressing either antisense of HSA-miR-6891-5p or scrambled control, and altered mRNA transcript levels were assessed using microarrays. (A) Principal component analysis (PCA) was performed in order to visualize sample clustering and assess the variation among biological replicates ( N = 3 experimental and 2 controls samples). Clear circles represent the centroid of the sample clusters, and the ellipse represents 2× the standard deviation in the x - and y -axes, respectively. The first principal component accounts for 76.5% of the variance within the dataset, while the second principal component accounts for 8.6% of the variance within the dataset. (B) Hierarchical clustering of samples based upon identified differentially expressed transcripts from microarray analysis.
    Figure Legend Snippet: Identification of potential miR-6891-5p targets . COX cells were transduced with lentiviruses expressing either antisense of HSA-miR-6891-5p or scrambled control, and altered mRNA transcript levels were assessed using microarrays. (A) Principal component analysis (PCA) was performed in order to visualize sample clustering and assess the variation among biological replicates ( N = 3 experimental and 2 controls samples). Clear circles represent the centroid of the sample clusters, and the ellipse represents 2× the standard deviation in the x - and y -axes, respectively. The first principal component accounts for 76.5% of the variance within the dataset, while the second principal component accounts for 8.6% of the variance within the dataset. (B) Hierarchical clustering of samples based upon identified differentially expressed transcripts from microarray analysis.

    Techniques Used: Transduction, Expressing, Standard Deviation, Microarray

    Validation of miR-6891-5p-mediated posttranscriptional regulation of immunoglobulin heavy chain alpha 1 and 2 ( IGHA1 and IGHA2 ) transcripts . (A) COX cells were transduced with lentiviral constructs expressing either the scrambled control or antisense sequence of miR-6891-5p. Cells were harvested after 48 h of transduction, total RNA was purified, and both IGHA1 and IGHA2 expression were analyzed by qPCR (ΔΔCt, standard error bars shown, n = 3). (B) COX cells (5 × 10 8 ) were transduced with lentiviral construct expressing either the scrambled control or antisense sequence of miR-6891-5p. After 120 h, media were collected and analyzed by ELISA using IgA antibody (standard error bars shown, n = 3). (C) Predicted binding site and heteroduplex formed between the wild-type (WT) 3′UTR of IGHA2 and miR-6891-5p. The heteroduplex formed with IGHA1 is identical to that shown. (D) Predicted binding site and heteroduplex formed between the mutated (Mut) 3′UTR sequence of IGHA2 and miR-6891-5p. (E) Either the wild-type (WT) or mutated (Mut) 3′UTR sequence of IGHA2 was cloned downstream of the luciferase reporter, creating two separate constructs. The wild-type or mutant luciferase constructs alone or together with either the miR-6891-5p expression construct (miR overexpression) or the antisense miR-6891-5p expression construct (miR inhibition) were transfected into HEK293T cells. Luciferase assay was performed 24 h after transfection (standard error bars shown, n = 3). All p -values shown are calculated using a t -test.
    Figure Legend Snippet: Validation of miR-6891-5p-mediated posttranscriptional regulation of immunoglobulin heavy chain alpha 1 and 2 ( IGHA1 and IGHA2 ) transcripts . (A) COX cells were transduced with lentiviral constructs expressing either the scrambled control or antisense sequence of miR-6891-5p. Cells were harvested after 48 h of transduction, total RNA was purified, and both IGHA1 and IGHA2 expression were analyzed by qPCR (ΔΔCt, standard error bars shown, n = 3). (B) COX cells (5 × 10 8 ) were transduced with lentiviral construct expressing either the scrambled control or antisense sequence of miR-6891-5p. After 120 h, media were collected and analyzed by ELISA using IgA antibody (standard error bars shown, n = 3). (C) Predicted binding site and heteroduplex formed between the wild-type (WT) 3′UTR of IGHA2 and miR-6891-5p. The heteroduplex formed with IGHA1 is identical to that shown. (D) Predicted binding site and heteroduplex formed between the mutated (Mut) 3′UTR sequence of IGHA2 and miR-6891-5p. (E) Either the wild-type (WT) or mutated (Mut) 3′UTR sequence of IGHA2 was cloned downstream of the luciferase reporter, creating two separate constructs. The wild-type or mutant luciferase constructs alone or together with either the miR-6891-5p expression construct (miR overexpression) or the antisense miR-6891-5p expression construct (miR inhibition) were transfected into HEK293T cells. Luciferase assay was performed 24 h after transfection (standard error bars shown, n = 3). All p -values shown are calculated using a t -test.

    Techniques Used: Transduction, Construct, Expressing, Sequencing, Purification, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Binding Assay, Clone Assay, Luciferase, Mutagenesis, Over Expression, Inhibition, Transfection

    22) Product Images from "Analysis of circulating-microRNA expression in lactating Holstein cows under summer heat stress"

    Article Title: Analysis of circulating-microRNA expression in lactating Holstein cows under summer heat stress

    Journal: bioRxiv

    doi: 10.1101/2020.03.18.996777

    Volcano plot showing differentially expressed miRNAs between NHS and HS using transformed normalized data. |Fold change| value ≥ 2 and P
    Figure Legend Snippet: Volcano plot showing differentially expressed miRNAs between NHS and HS using transformed normalized data. |Fold change| value ≥ 2 and P

    Techniques Used: Transformation Assay

    23) Product Images from "A Plasma Biomarker Panel of Four MicroRNAs for the Diagnosis of Prostate Cancer"

    Article Title: A Plasma Biomarker Panel of Four MicroRNAs for the Diagnosis of Prostate Cancer

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-24424-w

    Screening of plasma miRNA markers in pooled patient and control samples and validation in the discovery cohort. ( a) Scatter plot of 372 cancer-associated miRNAs in a screening cohort of pooled plasma samples from prostate cancer patients and healthy controls screened using a miScript miRNA PCR array. miRNAs found to be differentially expressed between the patient and control groups are shown in black, and unchanged miRNAs are shown in grey. A fold regulation cut-off of 2.5 was selected for the analysis. (b) Scatter plot showing the 11 differentially regulated miRNAs re-analysed by qRT-PCR in patient samples in the discovery cohort (N = 61). The mean fold regulation of each miRNA across the patient and control samples was taken into account and those that were below the 2 fold regulation cut-off were excluded from further analysis. The selected miRNAs are shown in colour. (c) Relative levels of miR-4289, miR-326, miR-152-3p and miR-98-5p analysed by qRT-PCR as in ( b ) in patients vs healthy controls. Statistically significant differences were assessed using a Mann-Whitney U test; p values are shown after Bonferroni correction for multiple testing. Each data point represents a plasma sample, the horizontal middle line in each data set represents the mean, and the limits of the vertical lines represent the standard deviation.
    Figure Legend Snippet: Screening of plasma miRNA markers in pooled patient and control samples and validation in the discovery cohort. ( a) Scatter plot of 372 cancer-associated miRNAs in a screening cohort of pooled plasma samples from prostate cancer patients and healthy controls screened using a miScript miRNA PCR array. miRNAs found to be differentially expressed between the patient and control groups are shown in black, and unchanged miRNAs are shown in grey. A fold regulation cut-off of 2.5 was selected for the analysis. (b) Scatter plot showing the 11 differentially regulated miRNAs re-analysed by qRT-PCR in patient samples in the discovery cohort (N = 61). The mean fold regulation of each miRNA across the patient and control samples was taken into account and those that were below the 2 fold regulation cut-off were excluded from further analysis. The selected miRNAs are shown in colour. (c) Relative levels of miR-4289, miR-326, miR-152-3p and miR-98-5p analysed by qRT-PCR as in ( b ) in patients vs healthy controls. Statistically significant differences were assessed using a Mann-Whitney U test; p values are shown after Bonferroni correction for multiple testing. Each data point represents a plasma sample, the horizontal middle line in each data set represents the mean, and the limits of the vertical lines represent the standard deviation.

    Techniques Used: Polymerase Chain Reaction, Quantitative RT-PCR, MANN-WHITNEY, Standard Deviation

    24) Product Images from "A Plasma Biomarker Panel of Four MicroRNAs for the Diagnosis of Prostate Cancer"

    Article Title: A Plasma Biomarker Panel of Four MicroRNAs for the Diagnosis of Prostate Cancer

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-24424-w

    Screening of plasma miRNA markers in pooled patient and control samples and validation in the discovery cohort. ( a) Scatter plot of 372 cancer-associated miRNAs in a screening cohort of pooled plasma samples from prostate cancer patients and healthy controls screened using a miScript miRNA PCR array. miRNAs found to be differentially expressed between the patient and control groups are shown in black, and unchanged miRNAs are shown in grey. A fold regulation cut-off of 2.5 was selected for the analysis. (b) Scatter plot showing the 11 differentially regulated miRNAs re-analysed by qRT-PCR in patient samples in the discovery cohort (N = 61). The mean fold regulation of each miRNA across the patient and control samples was taken into account and those that were below the 2 fold regulation cut-off were excluded from further analysis. The selected miRNAs are shown in colour. (c) Relative levels of miR-4289, miR-326, miR-152-3p and miR-98-5p analysed by qRT-PCR as in ( b ) in patients vs healthy controls. Statistically significant differences were assessed using a Mann-Whitney U test; p values are shown after Bonferroni correction for multiple testing. Each data point represents a plasma sample, the horizontal middle line in each data set represents the mean, and the limits of the vertical lines represent the standard deviation.
    Figure Legend Snippet: Screening of plasma miRNA markers in pooled patient and control samples and validation in the discovery cohort. ( a) Scatter plot of 372 cancer-associated miRNAs in a screening cohort of pooled plasma samples from prostate cancer patients and healthy controls screened using a miScript miRNA PCR array. miRNAs found to be differentially expressed between the patient and control groups are shown in black, and unchanged miRNAs are shown in grey. A fold regulation cut-off of 2.5 was selected for the analysis. (b) Scatter plot showing the 11 differentially regulated miRNAs re-analysed by qRT-PCR in patient samples in the discovery cohort (N = 61). The mean fold regulation of each miRNA across the patient and control samples was taken into account and those that were below the 2 fold regulation cut-off were excluded from further analysis. The selected miRNAs are shown in colour. (c) Relative levels of miR-4289, miR-326, miR-152-3p and miR-98-5p analysed by qRT-PCR as in ( b ) in patients vs healthy controls. Statistically significant differences were assessed using a Mann-Whitney U test; p values are shown after Bonferroni correction for multiple testing. Each data point represents a plasma sample, the horizontal middle line in each data set represents the mean, and the limits of the vertical lines represent the standard deviation.

    Techniques Used: Polymerase Chain Reaction, Quantitative RT-PCR, MANN-WHITNEY, Standard Deviation

    25) Product Images from "Retaining MKP1 expression and attenuating JNK-mediated apoptosis by RIP1 for cisplatin resistance through miR-940 inhibition"

    Article Title: Retaining MKP1 expression and attenuating JNK-mediated apoptosis by RIP1 for cisplatin resistance through miR-940 inhibition

    Journal: Oncotarget

    doi:

    Increased miR-940 expression is involved in MKP1 suppression in RIP1 knockdown cells (A) Total RNA isolated from A549 cells (control and RIP1 knockdown) was used for detection of miR-940 with qPCR. (B) The cells were transfected with negative control or miR-940 miScript miRNA inhibitor for 48 h, MKP1 expression was detected with Western blot, and β-actin was detected as an input control. The intensity of the individual bands was quantified by Quantity One® Software and normalized to the corresponding input control (β-actin) bands. (C) The cells were transfected with the indicated miRNA inhibitor (10 nM) for 24h, and treated with cisplatin (20 μM) for 8 h. JNK1 and phospho-JNK1 were detected with Western blot. β-actin was detected as an input control. (D) The cells transfected with negative control or miR-940 inhibitor (10nM) for 24h, then the cells were left untreated or treated with cisplatin (20 μM) for an additional 48 h. Cell death was detected with LDH assay. Data shown are the mean±SD. *p
    Figure Legend Snippet: Increased miR-940 expression is involved in MKP1 suppression in RIP1 knockdown cells (A) Total RNA isolated from A549 cells (control and RIP1 knockdown) was used for detection of miR-940 with qPCR. (B) The cells were transfected with negative control or miR-940 miScript miRNA inhibitor for 48 h, MKP1 expression was detected with Western blot, and β-actin was detected as an input control. The intensity of the individual bands was quantified by Quantity One® Software and normalized to the corresponding input control (β-actin) bands. (C) The cells were transfected with the indicated miRNA inhibitor (10 nM) for 24h, and treated with cisplatin (20 μM) for 8 h. JNK1 and phospho-JNK1 were detected with Western blot. β-actin was detected as an input control. (D) The cells transfected with negative control or miR-940 inhibitor (10nM) for 24h, then the cells were left untreated or treated with cisplatin (20 μM) for an additional 48 h. Cell death was detected with LDH assay. Data shown are the mean±SD. *p

    Techniques Used: Expressing, Isolation, Real-time Polymerase Chain Reaction, Transfection, Negative Control, Western Blot, Software, Lactate Dehydrogenase Assay

    26) Product Images from "A Panel of Plasma Exosomal miRNAs as Potential Biomarkers for Differential Diagnosis of Thyroid Nodules"

    Article Title: A Panel of Plasma Exosomal miRNAs as Potential Biomarkers for Differential Diagnosis of Thyroid Nodules

    Journal: Frontiers in Genetics

    doi: 10.3389/fgene.2020.00449

    Expression levels of the selected plasma exosomal miRNA candidates in the validation set. Plasma exosomal miRNA was isolated from 30 benign thyroid nodules, 35 PTC cases, and 31 healthy controls. A total of 4 ng of miRNA was reverse transcribed using a QIAGEN miScript II RT kit. After 1:10 dilution, 2 μL of cDNA template was used for quantification by real-time PCR. Plasma levels of exosomal (A) miR-16-2-3p ( p
    Figure Legend Snippet: Expression levels of the selected plasma exosomal miRNA candidates in the validation set. Plasma exosomal miRNA was isolated from 30 benign thyroid nodules, 35 PTC cases, and 31 healthy controls. A total of 4 ng of miRNA was reverse transcribed using a QIAGEN miScript II RT kit. After 1:10 dilution, 2 μL of cDNA template was used for quantification by real-time PCR. Plasma levels of exosomal (A) miR-16-2-3p ( p

    Techniques Used: Expressing, Isolation, Real-time Polymerase Chain Reaction

    27) Product Images from "Transcriptomic microRNA Profiling of Dendritic Cells in Response to Gut Microbiota-Secreted Vesicles"

    Article Title: Transcriptomic microRNA Profiling of Dendritic Cells in Response to Gut Microbiota-Secreted Vesicles

    Journal: Cells

    doi: 10.3390/cells9061534

    Correlation between expression levels of miR-155-5p, miR-146a-5p and miR-146b-5p and their targets at a short time post-stimulation. Immature mo-DCs were stimulated with EcN or ECOR12 MVs for 6 h. ( A ) Relative levels of the indicated miRNAs and ( B ) relative levels of the known target mRNAs, SOCS1, TAB2 (miR-155-5p) and TRAF (miR-146-5p), were assessed by RT-qPCR and normalized, as described in Figure 6 and Figure 7 ’s legends, respectively. Data are presented as fold changes compared to untreated control cells (four independent biological experiments which were performed in triplicate). Statistical differences were evaluated by one-way ANOVA, followed by Bonferroni’s test. * Significance against untreated control cells.
    Figure Legend Snippet: Correlation between expression levels of miR-155-5p, miR-146a-5p and miR-146b-5p and their targets at a short time post-stimulation. Immature mo-DCs were stimulated with EcN or ECOR12 MVs for 6 h. ( A ) Relative levels of the indicated miRNAs and ( B ) relative levels of the known target mRNAs, SOCS1, TAB2 (miR-155-5p) and TRAF (miR-146-5p), were assessed by RT-qPCR and normalized, as described in Figure 6 and Figure 7 ’s legends, respectively. Data are presented as fold changes compared to untreated control cells (four independent biological experiments which were performed in triplicate). Statistical differences were evaluated by one-way ANOVA, followed by Bonferroni’s test. * Significance against untreated control cells.

    Techniques Used: Expressing, Quantitative RT-PCR

    RT-qPCR validation of selected miRNAs . Immature monocyte-derived dendritic cells (mo-DCs) were challenged for 24 h with MVs (10 µg/mL) from EcN (white bars) or ECOR12 (gray bars). Untreated control DCs are shown as black bars. ( A ) Cycle-threshold values for the reference genes (RNU-6, miR-421 and miR-let7f-5p). ( B ) Relative miRNA levels were measured by RT-qPCR and normalized to the three indicated reference genes. Data (mean ± SEM) are from six independent biological experiments (six donors) performed in triplicate and are presented as fold changes compared to untreated control cells. Statistical differences were evaluated by one-way ANOVA, followed by Bonferroni’s test ( p
    Figure Legend Snippet: RT-qPCR validation of selected miRNAs . Immature monocyte-derived dendritic cells (mo-DCs) were challenged for 24 h with MVs (10 µg/mL) from EcN (white bars) or ECOR12 (gray bars). Untreated control DCs are shown as black bars. ( A ) Cycle-threshold values for the reference genes (RNU-6, miR-421 and miR-let7f-5p). ( B ) Relative miRNA levels were measured by RT-qPCR and normalized to the three indicated reference genes. Data (mean ± SEM) are from six independent biological experiments (six donors) performed in triplicate and are presented as fold changes compared to untreated control cells. Statistical differences were evaluated by one-way ANOVA, followed by Bonferroni’s test ( p

    Techniques Used: Quantitative RT-PCR, Derivative Assay

    28) Product Images from "Identification of a potential non-coding RNA biomarker signature for amyotrophic lateral sclerosis"

    Article Title: Identification of a potential non-coding RNA biomarker signature for amyotrophic lateral sclerosis

    Journal: Brain communications

    doi: 10.1093/braincomms/fcaa053

    Differential expression of ncRNA biomarkers in ALS patient serum samples using RT-qPCR in the BioMOx discovery cohort. ( A ) Overall effects of disease state were found across all six ncRNA. ( B ) A correlation (Pearson’s) was found between the progression of ALS as determined by the monthly change in the ALSFRS-R score and hsa-miR-16-5p and hsa-miR-92a-3p expression. hsa-miR-21-5p/hsa-piR-33151: one-way ANOVA with Tukey post-hoc; hsa-miR-16-5p/hsa-miR-92a-3p/TRV-AAC4-1.1/TRA-AGC6-1.1: Welch’s one-way ANOVA with Games–Howell post hoc . Normalized to hsa-miR-718 and hsa-piR-31068. Relative expression to the average expression of healthy controls. Healthy controls n = 21, disease mimics n = 16, slow-progressing ALS (ALS-SP) n = 23, fast-progressing ALS (ALS-FP) n = 21. Bars: average ± SD.
    Figure Legend Snippet: Differential expression of ncRNA biomarkers in ALS patient serum samples using RT-qPCR in the BioMOx discovery cohort. ( A ) Overall effects of disease state were found across all six ncRNA. ( B ) A correlation (Pearson’s) was found between the progression of ALS as determined by the monthly change in the ALSFRS-R score and hsa-miR-16-5p and hsa-miR-92a-3p expression. hsa-miR-21-5p/hsa-piR-33151: one-way ANOVA with Tukey post-hoc; hsa-miR-16-5p/hsa-miR-92a-3p/TRV-AAC4-1.1/TRA-AGC6-1.1: Welch’s one-way ANOVA with Games–Howell post hoc . Normalized to hsa-miR-718 and hsa-piR-31068. Relative expression to the average expression of healthy controls. Healthy controls n = 21, disease mimics n = 16, slow-progressing ALS (ALS-SP) n = 23, fast-progressing ALS (ALS-FP) n = 21. Bars: average ± SD.

    Techniques Used: Expressing, Quantitative RT-PCR

    29) Product Images from "miR-125a regulates cell cycle, proliferation, and apoptosis by targeting the ErbB Pathway in Acute Myeloid Leukemia"

    Article Title: miR-125a regulates cell cycle, proliferation, and apoptosis by targeting the ErbB Pathway in Acute Myeloid Leukemia

    Journal: Leukemia research

    doi: 10.1016/j.leukres.2013.12.021

    miR-125a is epigenetically silenced in AML
    Figure Legend Snippet: miR-125a is epigenetically silenced in AML

    Techniques Used:

    Restored miR-125a expression causes decreased cell proliferation, cell cycle progression, and enhanced apoptosis
    Figure Legend Snippet: Restored miR-125a expression causes decreased cell proliferation, cell cycle progression, and enhanced apoptosis

    Techniques Used: Expressing

    30) Product Images from "Revised annotation and characterization of novel Aedes albopictus miRNAs and their potential functions in dengue virus infection"

    Article Title: Revised annotation and characterization of novel Aedes albopictus miRNAs and their potential functions in dengue virus infection

    Journal: bioRxiv

    doi: 10.1101/2020.03.01.972398

    qRT-PCR of selected novel miRNAs Two-tailed t- test was used for all comparisons; *p
    Figure Legend Snippet: qRT-PCR of selected novel miRNAs Two-tailed t- test was used for all comparisons; *p

    Techniques Used: Quantitative RT-PCR, Two Tailed Test

    31) Product Images from "Leishmania (Leishmania) amazonensis induces macrophage miR-294 and miR-721 expression and modulates infection by targeting NOS2 and L-arginine metabolism"

    Article Title: Leishmania (Leishmania) amazonensis induces macrophage miR-294 and miR-721 expression and modulates infection by targeting NOS2 and L-arginine metabolism

    Journal: Scientific Reports

    doi: 10.1038/srep44141

    miR-294 and miR-721 bind to Nos2 mRNA and regulating NOS2 expression and L . ( L .) amazonensis infectivity. BMDMs (5 × 10 5 ) were plated into chamber slides overnight and transfected with negative control, 0.1 or 0.5 μM miScript Target Protector for miR-294/ Nos2 or miR-721/ Nos 2 or negative control for 24 h. Then, the BMDMs were co-cultivated with La -WT L . ( L .) amazonensis (MOI 5:1) for 4 h, and the cultures were washed. After 4 h and 24 h of infection, the samples were analyzed for NOS2 protein levels ( A , D ) via in-cell western, NO production ( B , E ) via flow cytometry DAF-FM fluorescence analysis, and for infectivity ( C , F ) via microscopy analysis, counting of the numbers of infected macrophages and amastigotes per macrophage (n = 1,000 macrophages/treatment). The values were normalized based on the average of untreated infected macrophages. Each bar represents the average ± SEM of the values obtained in 3 independent experiments (n = 4–6). Statistical significance was determined based on two-tailed Student’s t test. *p
    Figure Legend Snippet: miR-294 and miR-721 bind to Nos2 mRNA and regulating NOS2 expression and L . ( L .) amazonensis infectivity. BMDMs (5 × 10 5 ) were plated into chamber slides overnight and transfected with negative control, 0.1 or 0.5 μM miScript Target Protector for miR-294/ Nos2 or miR-721/ Nos 2 or negative control for 24 h. Then, the BMDMs were co-cultivated with La -WT L . ( L .) amazonensis (MOI 5:1) for 4 h, and the cultures were washed. After 4 h and 24 h of infection, the samples were analyzed for NOS2 protein levels ( A , D ) via in-cell western, NO production ( B , E ) via flow cytometry DAF-FM fluorescence analysis, and for infectivity ( C , F ) via microscopy analysis, counting of the numbers of infected macrophages and amastigotes per macrophage (n = 1,000 macrophages/treatment). The values were normalized based on the average of untreated infected macrophages. Each bar represents the average ± SEM of the values obtained in 3 independent experiments (n = 4–6). Statistical significance was determined based on two-tailed Student’s t test. *p

    Techniques Used: Expressing, Infection, Transfection, Negative Control, In-Cell ELISA, Flow Cytometry, Cytometry, Fluorescence, Microscopy, Two Tailed Test

    32) Product Images from "Ear atresia: Is there a role of apoptosis-regulating miRNAs?"

    Article Title: Ear atresia: Is there a role of apoptosis-regulating miRNAs?

    Journal: Northern Clinics of Istanbul

    doi: 10.14744/nci.2017.26680

    Clustergram of miRNA expressions in patients with ear atresia compared with controls. This clustergram was created with online QIAGEN miScript Primer Assay, Data Analysis Center. Accordingly, expression levels of 10 miRNAs were decreased (green) in the patients compared with the control group (red). The decreased expression levels of miR146a (p=0.000), miR222 (p=0.009), and miR126 (p=0.041) were statistically significant.
    Figure Legend Snippet: Clustergram of miRNA expressions in patients with ear atresia compared with controls. This clustergram was created with online QIAGEN miScript Primer Assay, Data Analysis Center. Accordingly, expression levels of 10 miRNAs were decreased (green) in the patients compared with the control group (red). The decreased expression levels of miR146a (p=0.000), miR222 (p=0.009), and miR126 (p=0.041) were statistically significant.

    Techniques Used: Expressing

    33) Product Images from "Arsenic Alters Exosome Quantity and Cargo to Mediate Stem Cell Recruitment Into a Cancer Stem Cell-Like Phenotype"

    Article Title: Arsenic Alters Exosome Quantity and Cargo to Mediate Stem Cell Recruitment Into a Cancer Stem Cell-Like Phenotype

    Journal: Toxicological Sciences

    doi: 10.1093/toxsci/kfy176

    Arsenite transformed CAsE-PE cells secrete more exosomes than RWPE-1 cells. A, Nanoparticle tracking analysis of isolated exosomes. B, Western blot analysis of exosomal markers CD9 and CD81 and nonexosomal markers GRP94, CANX, and HSP70. C, CAsE-PE cells secrete 702% more exosomes than RWPE-1 cells (1 factor ANOVA, p = .0002, n = 3, Bars ± SEM).
    Figure Legend Snippet: Arsenite transformed CAsE-PE cells secrete more exosomes than RWPE-1 cells. A, Nanoparticle tracking analysis of isolated exosomes. B, Western blot analysis of exosomal markers CD9 and CD81 and nonexosomal markers GRP94, CANX, and HSP70. C, CAsE-PE cells secrete 702% more exosomes than RWPE-1 cells (1 factor ANOVA, p = .0002, n = 3, Bars ± SEM).

    Techniques Used: Transformation Assay, Isolation, Western Blot

    A, Western blot analysis of KRAS protein levels in cell lysate and exosomes following KRAS KD. B, Western blot analysis of KRAS, BCL-XL, and BCL2 protein levels in SCs following 3 weeks of coculture with KRAS-KD CAsE-PE cells. C, Secreted MMP-2 (2 factor ANOVA, time ( p = .34), KD ( p = .23), interaction ( p = .34)), and (D) MMP-9 activity (2 factor ANOVA, time ( p = .38), KD ( p = .013), interaction ( p = .38)) in cocultured SCs. E, Invasive capacity (one factor ANOVA, p = .02) and (F) colony formation in soft agar (1 factor ANOVA, p = .8) of SCs after 3 weeks of coculture. Asterisk denotes statistical significance ( p
    Figure Legend Snippet: A, Western blot analysis of KRAS protein levels in cell lysate and exosomes following KRAS KD. B, Western blot analysis of KRAS, BCL-XL, and BCL2 protein levels in SCs following 3 weeks of coculture with KRAS-KD CAsE-PE cells. C, Secreted MMP-2 (2 factor ANOVA, time ( p = .34), KD ( p = .23), interaction ( p = .34)), and (D) MMP-9 activity (2 factor ANOVA, time ( p = .38), KD ( p = .013), interaction ( p = .38)) in cocultured SCs. E, Invasive capacity (one factor ANOVA, p = .02) and (F) colony formation in soft agar (1 factor ANOVA, p = .8) of SCs after 3 weeks of coculture. Asterisk denotes statistical significance ( p

    Techniques Used: Western Blot, Activity Assay

    34) Product Images from "CD177-mediated nanoparticle targeting of human and mouse neutrophils"

    Article Title: CD177-mediated nanoparticle targeting of human and mouse neutrophils

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0200444

    Mouse C5aR1 ASO results in knockdown of mouse C5aR1-GFP in CHO transfectants. CHO cells expressing mouse C5aR1-GFP were transfected with 50 nM or 100 nM LNA GapmeR ASO. The cells were analyzed for C5aR1-GFP expression and mRNA levels 72 h post transfection. A. Relative receptor knockdown was measured by flow cytometry. The percentage knockdown was calculated based on the number of cells to the left of the gate relative to the negative control ASO. B. Relative gene expression was calculated from quantification cycle (Cq) values obtained by RT-qPCR using the ΔΔCq method. To control for possible experimental variation, the qPCR was carried out using two sets of mouse C5aR1 primers (C5aR1 208–402 and 221–430), and two sets of reference primers. The results in the left panel show the relative quantity of C5aR1 mRNA normalized to Eif3i, and the results in the right panel show the relative quantity of C5aR1 mRNA normalized to Vezt. Mock transfected cells received no ASO and non-targeting control (NTC) cells were transfected with a non-targeting ASO. The RT-qPCR was carried out with triplicate samples ± SD. One-way analysis of variance at 95% confidence interval showed that the relative mRNA expression levels were significantly lower in the ASO treated cells compared to the mock transfected and non-targeting ASO cells ( p value
    Figure Legend Snippet: Mouse C5aR1 ASO results in knockdown of mouse C5aR1-GFP in CHO transfectants. CHO cells expressing mouse C5aR1-GFP were transfected with 50 nM or 100 nM LNA GapmeR ASO. The cells were analyzed for C5aR1-GFP expression and mRNA levels 72 h post transfection. A. Relative receptor knockdown was measured by flow cytometry. The percentage knockdown was calculated based on the number of cells to the left of the gate relative to the negative control ASO. B. Relative gene expression was calculated from quantification cycle (Cq) values obtained by RT-qPCR using the ΔΔCq method. To control for possible experimental variation, the qPCR was carried out using two sets of mouse C5aR1 primers (C5aR1 208–402 and 221–430), and two sets of reference primers. The results in the left panel show the relative quantity of C5aR1 mRNA normalized to Eif3i, and the results in the right panel show the relative quantity of C5aR1 mRNA normalized to Vezt. Mock transfected cells received no ASO and non-targeting control (NTC) cells were transfected with a non-targeting ASO. The RT-qPCR was carried out with triplicate samples ± SD. One-way analysis of variance at 95% confidence interval showed that the relative mRNA expression levels were significantly lower in the ASO treated cells compared to the mock transfected and non-targeting ASO cells ( p value

    Techniques Used: Allele-specific Oligonucleotide, Expressing, Transfection, Flow Cytometry, Cytometry, Negative Control, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

    Mouse C5aR1 siRNA and human C5aR1 siRNA pool result in receptor knockdown. CHO cells expressing mouse C5aR1-GFP were transfected with 100 nM mouse C5aR1 ON-TARGETplus SMART siRNA–6, 100 nM GFP siRNA (positive control), or 100 nM negative control siRNA. 72 h post transfection cells were analyzed by flow cytometry to measure the relative expression of mouse C5aR1-GFP (left panel). CHO cells expressing human C5aR1-GFP were transfected with 100 nM human C5aR1 ON-TARGETplus SMARTpool siRNA, 100 nM GFP siRNA (positive control), or 100 nM negative control siRNA (right panel). Relative knockdown is based on the percentage of the cells that are to the left of the gate relative to the negative control sample. The experiment was carried out twice with similar results.
    Figure Legend Snippet: Mouse C5aR1 siRNA and human C5aR1 siRNA pool result in receptor knockdown. CHO cells expressing mouse C5aR1-GFP were transfected with 100 nM mouse C5aR1 ON-TARGETplus SMART siRNA–6, 100 nM GFP siRNA (positive control), or 100 nM negative control siRNA. 72 h post transfection cells were analyzed by flow cytometry to measure the relative expression of mouse C5aR1-GFP (left panel). CHO cells expressing human C5aR1-GFP were transfected with 100 nM human C5aR1 ON-TARGETplus SMARTpool siRNA, 100 nM GFP siRNA (positive control), or 100 nM negative control siRNA (right panel). Relative knockdown is based on the percentage of the cells that are to the left of the gate relative to the negative control sample. The experiment was carried out twice with similar results.

    Techniques Used: Expressing, Transfection, Positive Control, Negative Control, Flow Cytometry, Cytometry

    35) Product Images from "A Novel S100A8/A9 Induced Fingerprint of Mesenchymal Stem Cells associated with Enhanced Wound Healing"

    Article Title: A Novel S100A8/A9 Induced Fingerprint of Mesenchymal Stem Cells associated with Enhanced Wound Healing

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-24425-9

    Validation of RNA-seq data by qRT-PCR. Total RNA and miRNA was extracted from control MSCs and from MSCs which have been treated with 0.5 µg/mL S100A8/A9. Isolated RNAs were assessed for quality with Bioanalyzer and reverse-transcribed using the Reverse Transcription System. The primers targeted (a) CCR7, (b) IL-32, (c) ICOSLG, (d) PRSS36, (e) SERPINA9, (f) PI15, ( g ) MMP27, (h) SPOCK2, ( i ) DSC2, ( j ) hsa-miR-582-5p, to amplify cDNA using 2x power SYBR Green Mix (Applied Biosystems). Relative mRNA or miR levels were calculated by normalizing to β-actin and RNU6B in case of miRNA. Box plots are expressed as mean with min-max wiskers representing the normalized expression level of each gene of MSCs treated with S100A8/A9 as opposed to control MSCs. Data were analyzed using the unpaired, two-tailed non-parametric Student’s t test using Wilcoxon signed rank-test and the differential expression of the qPCR results were expressed as log 2 fold change with the respective p-values. P-value
    Figure Legend Snippet: Validation of RNA-seq data by qRT-PCR. Total RNA and miRNA was extracted from control MSCs and from MSCs which have been treated with 0.5 µg/mL S100A8/A9. Isolated RNAs were assessed for quality with Bioanalyzer and reverse-transcribed using the Reverse Transcription System. The primers targeted (a) CCR7, (b) IL-32, (c) ICOSLG, (d) PRSS36, (e) SERPINA9, (f) PI15, ( g ) MMP27, (h) SPOCK2, ( i ) DSC2, ( j ) hsa-miR-582-5p, to amplify cDNA using 2x power SYBR Green Mix (Applied Biosystems). Relative mRNA or miR levels were calculated by normalizing to β-actin and RNU6B in case of miRNA. Box plots are expressed as mean with min-max wiskers representing the normalized expression level of each gene of MSCs treated with S100A8/A9 as opposed to control MSCs. Data were analyzed using the unpaired, two-tailed non-parametric Student’s t test using Wilcoxon signed rank-test and the differential expression of the qPCR results were expressed as log 2 fold change with the respective p-values. P-value

    Techniques Used: RNA Sequencing Assay, Quantitative RT-PCR, Isolation, SYBR Green Assay, Expressing, Two Tailed Test, Real-time Polymerase Chain Reaction

    36) Product Images from "MicroRNA-210 suppresses glucocorticoid receptor expression in response to hypoxia in fetal rat cardiomyocytes"

    Article Title: MicroRNA-210 suppresses glucocorticoid receptor expression in response to hypoxia in fetal rat cardiomyocytes

    Journal: Oncotarget

    doi: 10.18632/oncotarget.17801

    Hypoxia induces miR-210 expression and suppresses GR expression in fetal rat hearts ( A ) Hearts were isolated from E21 fetuses from pregnant rats treated with normoxia or hypoxia at 10.5% O 2 from day 15 to day 21 of gestation, and miR-210 expression was measured by miScript miR real-time RT-qPCR. n = 8. ( B , C , D ) Cardiomyocytes isolated from E21 fetuses were treated with normoxia or hypoxia (1% O 2 ) for 24 hours. MiR-210 expression was measured by miScript miR real-time qRT-PCR (B), n = 6; GR protein abundance was measured by Western blot (C), n = 4–5, and GR mRNA abundance was determined by real-time qRT-PCR (D), n = 6. Data are mean ± SEM. * p
    Figure Legend Snippet: Hypoxia induces miR-210 expression and suppresses GR expression in fetal rat hearts ( A ) Hearts were isolated from E21 fetuses from pregnant rats treated with normoxia or hypoxia at 10.5% O 2 from day 15 to day 21 of gestation, and miR-210 expression was measured by miScript miR real-time RT-qPCR. n = 8. ( B , C , D ) Cardiomyocytes isolated from E21 fetuses were treated with normoxia or hypoxia (1% O 2 ) for 24 hours. MiR-210 expression was measured by miScript miR real-time qRT-PCR (B), n = 6; GR protein abundance was measured by Western blot (C), n = 4–5, and GR mRNA abundance was determined by real-time qRT-PCR (D), n = 6. Data are mean ± SEM. * p

    Techniques Used: Expressing, Isolation, Quantitative RT-PCR, Western Blot

    37) Product Images from "A Plasma Biomarker Panel of Four MicroRNAs for the Diagnosis of Prostate Cancer"

    Article Title: A Plasma Biomarker Panel of Four MicroRNAs for the Diagnosis of Prostate Cancer

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-24424-w

    Screening of plasma miRNA markers in pooled patient and control samples and validation in the discovery cohort. ( a) Scatter plot of 372 cancer-associated miRNAs in a screening cohort of pooled plasma samples from prostate cancer patients and healthy controls screened using a miScript miRNA PCR array. miRNAs found to be differentially expressed between the patient and control groups are shown in black, and unchanged miRNAs are shown in grey. A fold regulation cut-off of 2.5 was selected for the analysis. (b) Scatter plot showing the 11 differentially regulated miRNAs re-analysed by qRT-PCR in patient samples in the discovery cohort (N = 61). The mean fold regulation of each miRNA across the patient and control samples was taken into account and those that were below the 2 fold regulation cut-off were excluded from further analysis. The selected miRNAs are shown in colour. (c) Relative levels of miR-4289, miR-326, miR-152-3p and miR-98-5p analysed by qRT-PCR as in ( b ) in patients vs healthy controls. Statistically significant differences were assessed using a Mann-Whitney U test; p values are shown after Bonferroni correction for multiple testing. Each data point represents a plasma sample, the horizontal middle line in each data set represents the mean, and the limits of the vertical lines represent the standard deviation.
    Figure Legend Snippet: Screening of plasma miRNA markers in pooled patient and control samples and validation in the discovery cohort. ( a) Scatter plot of 372 cancer-associated miRNAs in a screening cohort of pooled plasma samples from prostate cancer patients and healthy controls screened using a miScript miRNA PCR array. miRNAs found to be differentially expressed between the patient and control groups are shown in black, and unchanged miRNAs are shown in grey. A fold regulation cut-off of 2.5 was selected for the analysis. (b) Scatter plot showing the 11 differentially regulated miRNAs re-analysed by qRT-PCR in patient samples in the discovery cohort (N = 61). The mean fold regulation of each miRNA across the patient and control samples was taken into account and those that were below the 2 fold regulation cut-off were excluded from further analysis. The selected miRNAs are shown in colour. (c) Relative levels of miR-4289, miR-326, miR-152-3p and miR-98-5p analysed by qRT-PCR as in ( b ) in patients vs healthy controls. Statistically significant differences were assessed using a Mann-Whitney U test; p values are shown after Bonferroni correction for multiple testing. Each data point represents a plasma sample, the horizontal middle line in each data set represents the mean, and the limits of the vertical lines represent the standard deviation.

    Techniques Used: Polymerase Chain Reaction, Quantitative RT-PCR, MANN-WHITNEY, Standard Deviation

    38) Product Images from "miR-214-Dependent Increase of PHLPP2 Levels Mediates the Impairment of Insulin-Stimulated Akt Activation in Mouse Aortic Endothelial Cells Exposed to Methylglyoxal"

    Article Title: miR-214-Dependent Increase of PHLPP2 Levels Mediates the Impairment of Insulin-Stimulated Akt Activation in Mouse Aortic Endothelial Cells Exposed to Methylglyoxal

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19020522

    Effect of miR-214 on PHLPP2 regulation. ( a ) MAECs were transfected with a non-targeting control oligonucleotide (Ctr M; white bars) or miR-214 mimic (miR-214 M 50 nmol/L; black bars). Where indicated, MAECs were co-transfected with miScript miR-214 Target Protector (miR-214 TP 0.1 nmol/L). Protein lysates obtained from 48 h transfected MAECs were analyzed by Western blot with anti-PHLPP2 and anti-α-tubulin antibodies. Exposure timing of blots was of 2 min. Protein levels were quantified by the densitometric analysis of at least three independent experiments. Bars in the graphs represent the mean ± SD of the percent (%) over control (Ctr M). ( b ) MAECs were transfected for 24 h with non-targeting control oligonucleotide (Ctr M 50 nmol/L; white bars) or miR-214 mimic (miR-214 M 50 nmol/L; black bars) in the presence of the pEZX-MT06 control reporter vector or the pEZX-MT06/PHLPP2-3′UTR reporter vector. Firefly and Renilla luciferase activities were determined in cell lysates under each experimental condition. Results are normalized to Renilla activity. Bars in the graphs represent the mean ± SD of the fold over control (Ctr M). Statistical analysis was evaluated using Student’s t -test; *** p ≤ 0.001.
    Figure Legend Snippet: Effect of miR-214 on PHLPP2 regulation. ( a ) MAECs were transfected with a non-targeting control oligonucleotide (Ctr M; white bars) or miR-214 mimic (miR-214 M 50 nmol/L; black bars). Where indicated, MAECs were co-transfected with miScript miR-214 Target Protector (miR-214 TP 0.1 nmol/L). Protein lysates obtained from 48 h transfected MAECs were analyzed by Western blot with anti-PHLPP2 and anti-α-tubulin antibodies. Exposure timing of blots was of 2 min. Protein levels were quantified by the densitometric analysis of at least three independent experiments. Bars in the graphs represent the mean ± SD of the percent (%) over control (Ctr M). ( b ) MAECs were transfected for 24 h with non-targeting control oligonucleotide (Ctr M 50 nmol/L; white bars) or miR-214 mimic (miR-214 M 50 nmol/L; black bars) in the presence of the pEZX-MT06 control reporter vector or the pEZX-MT06/PHLPP2-3′UTR reporter vector. Firefly and Renilla luciferase activities were determined in cell lysates under each experimental condition. Results are normalized to Renilla activity. Bars in the graphs represent the mean ± SD of the fold over control (Ctr M). Statistical analysis was evaluated using Student’s t -test; *** p ≤ 0.001.

    Techniques Used: Transfection, Western Blot, Plasmid Preparation, Luciferase, Activity Assay

    39) Product Images from "Differential expression of microRNAs in retinal vasculopathy caused by selective Müller cell disruption"

    Article Title: Differential expression of microRNAs in retinal vasculopathy caused by selective Müller cell disruption

    Journal: Scientific Reports

    doi: 10.1038/srep28993

    Differentially expressed miRNAs 3 months after Müller cell disruption and its target gene validation with qRT-PCR. ( A ) Volcano plot showing differentially expressed miRNAs 3 months after selective Müller cell disruption. Red circles represent upregulated miRNAs and green circles indicate downregulated miRNAs. Vertical grey lines indicate fold changes, with a cut off ±2. The horizontal blue line represents a p-value of 0.05. n = 6 in each group. ( B ) qRT-PCR analysis of genes targeted by miR-200b after selective Müller cell disruption. qRT-PCR was conducted using retinas collected 3 months after induced Müller cell disruption. The boxes represent the interquartile range. The dotted lines within the boxes represent medians of gene expression. The whiskers indicate the maximum and minimum values of gene expression. *P
    Figure Legend Snippet: Differentially expressed miRNAs 3 months after Müller cell disruption and its target gene validation with qRT-PCR. ( A ) Volcano plot showing differentially expressed miRNAs 3 months after selective Müller cell disruption. Red circles represent upregulated miRNAs and green circles indicate downregulated miRNAs. Vertical grey lines indicate fold changes, with a cut off ±2. The horizontal blue line represents a p-value of 0.05. n = 6 in each group. ( B ) qRT-PCR analysis of genes targeted by miR-200b after selective Müller cell disruption. qRT-PCR was conducted using retinas collected 3 months after induced Müller cell disruption. The boxes represent the interquartile range. The dotted lines within the boxes represent medians of gene expression. The whiskers indicate the maximum and minimum values of gene expression. *P

    Techniques Used: Quantitative RT-PCR, Expressing

    40) Product Images from "Ex vivo miRNome analysis in Ptch1+/− cerebellum granule cells reveals a subset of miRNAs involved in radiation-induced medulloblastoma"

    Article Title: Ex vivo miRNome analysis in Ptch1+/− cerebellum granule cells reveals a subset of miRNAs involved in radiation-induced medulloblastoma

    Journal: Oncotarget

    doi: 10.18632/oncotarget.11938

    Real time PCR validation of selected statistically significant miRNAs from NGS and arrays on tumors Representative histological images of spontaneous ( A ) and radio-induced MB ( B ). ( C ) miRNAs expression in radio-induced MB with respect to the spontaneous one. Each data represent the mean and the standard deviation of three biological replicates. Student's t-test has been performed to evaluate the statistical significance of the comparisons. * P ≤ 0.05; ** P ≤ 0.005.
    Figure Legend Snippet: Real time PCR validation of selected statistically significant miRNAs from NGS and arrays on tumors Representative histological images of spontaneous ( A ) and radio-induced MB ( B ). ( C ) miRNAs expression in radio-induced MB with respect to the spontaneous one. Each data represent the mean and the standard deviation of three biological replicates. Student's t-test has been performed to evaluate the statistical significance of the comparisons. * P ≤ 0.05; ** P ≤ 0.005.

    Techniques Used: Real-time Polymerase Chain Reaction, Next-Generation Sequencing, Expressing, Standard Deviation

    Real time PCR validation on GCPs of selected statistically significant miRNAs from NGS and arrays Each data represent the mean and the standard deviation of three biological replicates with respect to the unirradiated WT GCPs. Student's t-test has been performed to calculate the statistical significance of comparisons. * P ≤ 0.05; ** P ≤ 0.005; *** P ≤ 0.0001.
    Figure Legend Snippet: Real time PCR validation on GCPs of selected statistically significant miRNAs from NGS and arrays Each data represent the mean and the standard deviation of three biological replicates with respect to the unirradiated WT GCPs. Student's t-test has been performed to calculate the statistical significance of comparisons. * P ≤ 0.05; ** P ≤ 0.005; *** P ≤ 0.0001.

    Techniques Used: Real-time Polymerase Chain Reaction, Next-Generation Sequencing, Standard Deviation

    Related Articles

    Real-time Polymerase Chain Reaction:

    Article Title: Gastrin-induced miR-222 promotes gastric tumor development by suppressing p27kip1
    Article Snippet: .. Eluted RNA was reverse transcribed into cDNA using the miScript RT II Kit (Qiagen, Sussex, UK) according to the manufacturer's procedures handbook and stored as undiluted cDNA at −20°C prior to real-time PCR. .. MicroRNA PCR array cDNA was prepared using the miScript SYBR Green PCR Kit (Qiagen, Sussex, UK) according to the manufacturer's instructions.

    Transfection:

    Article Title: MicroRNA-210 suppresses glucocorticoid receptor expression in response to hypoxia in fetal rat cardiomyocytes
    Article Snippet: .. Materials M199 medium was obtained from Hyclone (Logan, UT); MiRNA-210 mimic, HiPerFect transfection reagent, miScript II RT kit, miScript Primer Assay and miScript SYBR Green PCR kit were purchased from Qiagen (Valencia, CA). .. MiR-210 locked nucleic acid (miR-210-LNA) were purchased from Exiqon (Woburn, MA).

    Synthesized:

    Article Title: miR-34c-5p and CaMKII are involved in aldosterone-induced fibrosis in kidney collecting duct cells
    Article Snippet: .. mpkCCDc14 cells were treated with aldosterone (10−6 M; 3 or 5 days) or TGF-β (5 or 10 ng/ml; 3 days), and RNA was prepared by the mirVana miRNA Isolation Kit (Ambion; Thermo Fisher Scientific), according to the manufacturer’s instruction. cDNAs were synthesized using the miScript II RT Kit (Qiagen, Germantown, MD), as per the manufacturer’s protocol. .. Total RNA (1 μg), isolated from vehicle- or aldosterone-treated cells, was subjected to cDNA synthesis.

    Isolation:

    Article Title: miR-34c-5p and CaMKII are involved in aldosterone-induced fibrosis in kidney collecting duct cells
    Article Snippet: .. mpkCCDc14 cells were treated with aldosterone (10−6 M; 3 or 5 days) or TGF-β (5 or 10 ng/ml; 3 days), and RNA was prepared by the mirVana miRNA Isolation Kit (Ambion; Thermo Fisher Scientific), according to the manufacturer’s instruction. cDNAs were synthesized using the miScript II RT Kit (Qiagen, Germantown, MD), as per the manufacturer’s protocol. .. Total RNA (1 μg), isolated from vehicle- or aldosterone-treated cells, was subjected to cDNA synthesis.

    SYBR Green Assay:

    Article Title: Original Research: Stable expression of miR-34a mediates fetal hemoglobin induction in K562 cells
    Article Snippet: .. To quantify mature miR-34a levels, the miScript II RT and SYBR® Green PCR kit were used (Qiagen) per the manufacturer’s instructions. .. To generate miRNA cDNA, 500 ng of total RNA, HiSpec Buffer, Nucleic Mix, and miScript reverse transcriptase mix were incubated at 37℃ for 60 min. qPCR analysis was conducted with diluted cDNA template, universal primer, and miRNA specific primer per reaction; miR-34a levels were quantified using the 2−ΔΔCt method.

    Article Title: MicroRNA-210 suppresses glucocorticoid receptor expression in response to hypoxia in fetal rat cardiomyocytes
    Article Snippet: .. Materials M199 medium was obtained from Hyclone (Logan, UT); MiRNA-210 mimic, HiPerFect transfection reagent, miScript II RT kit, miScript Primer Assay and miScript SYBR Green PCR kit were purchased from Qiagen (Valencia, CA). .. MiR-210 locked nucleic acid (miR-210-LNA) were purchased from Exiqon (Woburn, MA).

    Expressing:

    Article Title: Epigenetic alterations in hippocampus of SAMP8 senescent mice and modulation by voluntary physical exercise
    Article Snippet: .. microRNA expression array RNA samples from 16 female individuals (four from each group: sedentary SAMR1, runner SAMR1, sedentary SAMP8, runner SAMP8) were converted to cDNA through a reverse transcription reaction using miScript II RT Kit (Qiagen, Hilden Germany) according to the manufacturer's instructions. .. The expression of 84 mature miRNAs was then analyzed using the miScript® miRNA PCR Array-Neurological Development and Disease miRNA PCR Array (Qiagen). miRNAs expression was measured in an ABI Prism 7900HT through SYBR-green-based real time PCR.

    Polymerase Chain Reaction:

    Article Title: Original Research: Stable expression of miR-34a mediates fetal hemoglobin induction in K562 cells
    Article Snippet: .. To quantify mature miR-34a levels, the miScript II RT and SYBR® Green PCR kit were used (Qiagen) per the manufacturer’s instructions. .. To generate miRNA cDNA, 500 ng of total RNA, HiSpec Buffer, Nucleic Mix, and miScript reverse transcriptase mix were incubated at 37℃ for 60 min. qPCR analysis was conducted with diluted cDNA template, universal primer, and miRNA specific primer per reaction; miR-34a levels were quantified using the 2−ΔΔCt method.

    Article Title: MicroRNA-210 suppresses glucocorticoid receptor expression in response to hypoxia in fetal rat cardiomyocytes
    Article Snippet: .. Materials M199 medium was obtained from Hyclone (Logan, UT); MiRNA-210 mimic, HiPerFect transfection reagent, miScript II RT kit, miScript Primer Assay and miScript SYBR Green PCR kit were purchased from Qiagen (Valencia, CA). .. MiR-210 locked nucleic acid (miR-210-LNA) were purchased from Exiqon (Woburn, MA).

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    Qiagen miscript ii rt
    miR-34a overexpression mediates HbF induction in K562 stable pools. K562 cells were transduced with the SMARTchoice™ shMIMIC miR-34a or scrambled lentivirus particles. After puromycin selection, cells were harvested at day 9 and day 16 for analysis (see Materials and methods) (a) Mature miR-34a expression was measured by RT-qPCR using the Qiagen’s <t>miScript</t> Primer Assay System. The ratio of miR-34a to RNU6 (housekeeping control) was plotted. The data were normalized to scramble control and reported as fold change of the mean ± standard error of the mean; * p
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    miR-34a overexpression mediates HbF induction in K562 stable pools. K562 cells were transduced with the SMARTchoice™ shMIMIC miR-34a or scrambled lentivirus particles. After puromycin selection, cells were harvested at day 9 and day 16 for analysis (see Materials and methods) (a) Mature miR-34a expression was measured by RT-qPCR using the Qiagen’s miScript Primer Assay System. The ratio of miR-34a to RNU6 (housekeeping control) was plotted. The data were normalized to scramble control and reported as fold change of the mean ± standard error of the mean; * p

    Journal: Experimental Biology and Medicine

    Article Title: Original Research: Stable expression of miR-34a mediates fetal hemoglobin induction in K562 cells

    doi: 10.1177/1535370216636725

    Figure Lengend Snippet: miR-34a overexpression mediates HbF induction in K562 stable pools. K562 cells were transduced with the SMARTchoice™ shMIMIC miR-34a or scrambled lentivirus particles. After puromycin selection, cells were harvested at day 9 and day 16 for analysis (see Materials and methods) (a) Mature miR-34a expression was measured by RT-qPCR using the Qiagen’s miScript Primer Assay System. The ratio of miR-34a to RNU6 (housekeeping control) was plotted. The data were normalized to scramble control and reported as fold change of the mean ± standard error of the mean; * p

    Article Snippet: To quantify mature miR-34a levels, the miScript II RT and SYBR® Green PCR kit were used (Qiagen) per the manufacturer’s instructions.

    Techniques: Over Expression, Transduction, Selection, Expressing, Quantitative RT-PCR

    Hypoxia induces miR-210 expression and suppresses GR expression in fetal rat hearts ( A ) Hearts were isolated from E21 fetuses from pregnant rats treated with normoxia or hypoxia at 10.5% O 2 from day 15 to day 21 of gestation, and miR-210 expression was measured by miScript miR real-time RT-qPCR. n = 8. ( B , C , D ) Cardiomyocytes isolated from E21 fetuses were treated with normoxia or hypoxia (1% O 2 ) for 24 hours. MiR-210 expression was measured by miScript miR real-time qRT-PCR (B), n = 6; GR protein abundance was measured by Western blot (C), n = 4–5, and GR mRNA abundance was determined by real-time qRT-PCR (D), n = 6. Data are mean ± SEM. * p

    Journal: Oncotarget

    Article Title: MicroRNA-210 suppresses glucocorticoid receptor expression in response to hypoxia in fetal rat cardiomyocytes

    doi: 10.18632/oncotarget.17801

    Figure Lengend Snippet: Hypoxia induces miR-210 expression and suppresses GR expression in fetal rat hearts ( A ) Hearts were isolated from E21 fetuses from pregnant rats treated with normoxia or hypoxia at 10.5% O 2 from day 15 to day 21 of gestation, and miR-210 expression was measured by miScript miR real-time RT-qPCR. n = 8. ( B , C , D ) Cardiomyocytes isolated from E21 fetuses were treated with normoxia or hypoxia (1% O 2 ) for 24 hours. MiR-210 expression was measured by miScript miR real-time qRT-PCR (B), n = 6; GR protein abundance was measured by Western blot (C), n = 4–5, and GR mRNA abundance was determined by real-time qRT-PCR (D), n = 6. Data are mean ± SEM. * p

    Article Snippet: Materials M199 medium was obtained from Hyclone (Logan, UT); MiRNA-210 mimic, HiPerFect transfection reagent, miScript II RT kit, miScript Primer Assay and miScript SYBR Green PCR kit were purchased from Qiagen (Valencia, CA).

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

    Validation of miR-196a expression in human skin fibroblasts ( a ) and human brain samples ( b ) by qRT-PCR. miR-196a expression was validated in human skin fibroblasts and human brain samples (from equivalent brain regions) using Qiagen miScript primer assays by qRT-PCR. The values were normalized to SNORD61 and presented as fold change. As represented by figure, miR-196a expression was downregulated in cALD fibroblasts and brain samples and upregulated in AMN fibroblasts and brain samples. B3 is brain sample from AMN that subsequently developed CNS disease. Downregulation of miR-196a expression in brain sample B3 indicates of cerebral involvement. Thirty-five percent of AMN patients develop cerebral involvement

    Journal: Molecular neurobiology

    Article Title: MicroRNA Profiling Identifies miR-196a as Differentially Expressed in Childhood Adrenoleukodystrophy and Adult Adrenomyeloneuropathy

    doi: 10.1007/s12035-016-9746-0

    Figure Lengend Snippet: Validation of miR-196a expression in human skin fibroblasts ( a ) and human brain samples ( b ) by qRT-PCR. miR-196a expression was validated in human skin fibroblasts and human brain samples (from equivalent brain regions) using Qiagen miScript primer assays by qRT-PCR. The values were normalized to SNORD61 and presented as fold change. As represented by figure, miR-196a expression was downregulated in cALD fibroblasts and brain samples and upregulated in AMN fibroblasts and brain samples. B3 is brain sample from AMN that subsequently developed CNS disease. Downregulation of miR-196a expression in brain sample B3 indicates of cerebral involvement. Thirty-five percent of AMN patients develop cerebral involvement

    Article Snippet: Total RNA that contains miRNA (600 ng) was used for cDNA synthesis using miScript II RT Kit (Qiagen) using miScript Reverse Transcriptase Mix, 10× miScript Nucleics Mix, and 5× miScript HiSpec Buffer.

    Techniques: Expressing, Quantitative RT-PCR

    Roles of miR-196a in target gene expression. To establish the regulation of target genes by miR-196a, miR-196a mimic was transfected in ABCD1 knockdown U87 astrocytes cells (U87 Lenti) and transfection efficiency evaluated by miScript qRT-PCR. SNORD61 was used as an internal control and represented as miR-196a/SNORD61 ( a ). The expression of target genes IKKα and IKKβ was analysed after the transfection of miR-196a mimic and normalized to GAPDH. The expression of these genes was significantly increased in ABCD1 silenced cells as compared to wild type but significantly decreased after transfection of mimic in U87 Lenti cells ( b and c ). ** P

    Journal: Molecular neurobiology

    Article Title: MicroRNA Profiling Identifies miR-196a as Differentially Expressed in Childhood Adrenoleukodystrophy and Adult Adrenomyeloneuropathy

    doi: 10.1007/s12035-016-9746-0

    Figure Lengend Snippet: Roles of miR-196a in target gene expression. To establish the regulation of target genes by miR-196a, miR-196a mimic was transfected in ABCD1 knockdown U87 astrocytes cells (U87 Lenti) and transfection efficiency evaluated by miScript qRT-PCR. SNORD61 was used as an internal control and represented as miR-196a/SNORD61 ( a ). The expression of target genes IKKα and IKKβ was analysed after the transfection of miR-196a mimic and normalized to GAPDH. The expression of these genes was significantly increased in ABCD1 silenced cells as compared to wild type but significantly decreased after transfection of mimic in U87 Lenti cells ( b and c ). ** P

    Article Snippet: Total RNA that contains miRNA (600 ng) was used for cDNA synthesis using miScript II RT Kit (Qiagen) using miScript Reverse Transcriptase Mix, 10× miScript Nucleics Mix, and 5× miScript HiSpec Buffer.

    Techniques: Expressing, Transfection, Quantitative RT-PCR

    Human cell differentiation and development miScript miRNA PCR array profiles. Group clustergram analysis of miRNA PCR array profiles analysis of differentially regulated miRNAs identified in control (undifferentiated hNSCs) and hNSCs seeded on 2D and 3D substrates and differentiated for 1W and 3W.

    Journal: Stem Cell Research & Therapy

    Article Title: The effects of microRNAs on human neural stem cell differentiation in two- and three-dimensional cultures

    doi: 10.1186/scrt437

    Figure Lengend Snippet: Human cell differentiation and development miScript miRNA PCR array profiles. Group clustergram analysis of miRNA PCR array profiles analysis of differentially regulated miRNAs identified in control (undifferentiated hNSCs) and hNSCs seeded on 2D and 3D substrates and differentiated for 1W and 3W.

    Article Snippet: For each array, a minimum of 250 ng total RNA was retrotranscribed by using miScript II RT Kit (Qiagen) according to the manufacturer’s instruction.

    Techniques: Cell Differentiation, Polymerase Chain Reaction