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mirna array assay  (LC Sciences)

 
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    LC Sciences mirna array assay
    TRPM7 negatively regulates miR-28-5p in glioma cells. (A) Total RNA from biological triplicate samples, either with scramble control (siCtrl, A423, A424, and A425) or siTRPM7 (A427, A428, and A434), were extracted and then parallelly subjected to <t>human</t> <t>miRNA</t> array assay. In-depth data analysis from miRNA microarray data resulted in a list of 16 downregulated and 10 upregulated miRNAs whose transcripts are statistically significant with fold changes greater than 2 by TRPM7 knock-down. (B,C) Validation of miRNA array by siTRPM7 by qPCR. miR-26b-5p, miR-4530, and miR-28-5p were validated by real-time miRNA quantification using stem-loop RT-PCR in glioma cell lines A172 (B) and SNB19 (C) . A set of endogenous RNAs SNOD48, SNOD44, SNOD47, and U6 served as endogenous controls in real-time miRNA quantification to normalize miRNA gene expression. (D) Validation of miRNA array by overexpressing TRPM7. The constructs of wt hTRPM7 (M7-WT), Δ kinase (M7-DK), K1648R (MT-KR), and control (Ctrl) were introduced into and expressed in glioma cell lines A172, U373MG, SNB19, and U87MG for 48 h. Transfection efficiency was determined by TRPM7 mRNA expression detected by qRT-PCR. GAPDH was used as an internal control to monitor the presence of amplified cDNA in samples (upper left). Western blots by overexpression of individual TRPM7 constructs confirmed the data from qRT-PCR, which showed significantly increased TRPM7 in M7-WT, M7-KR, and M7-DK compared to the control ( D , right). The levels of miR-28-5p were determined by stem-loop RT-PCR (lower left). All the data were represented as fold changes relative to the appropriate controls and are the mean ± SD of triplicated samples performed in three independent experiments. (* P < 0.05, ** P < 0.01).
    Mirna Array Assay, supplied by LC Sciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcr-based+mirna+microarray+platform/mirna+microarray/pmc06928690-52-18-23
    Average 90 stars, based on 1 article reviews
    mirna array assay - by Bioz Stars, 2026-11
    90/100 stars

    Images

    1) Product Images from "TRPM7 Induces Mechanistic Target of Rap1b Through the Downregulation of miR-28-5p in Glioma Proliferation and Invasion"

    Article Title: TRPM7 Induces Mechanistic Target of Rap1b Through the Downregulation of miR-28-5p in Glioma Proliferation and Invasion

    Journal: Frontiers in Oncology

    doi: 10.3389/fonc.2019.01413

    TRPM7 negatively regulates miR-28-5p in glioma cells. (A) Total RNA from biological triplicate samples, either with scramble control (siCtrl, A423, A424, and A425) or siTRPM7 (A427, A428, and A434), were extracted and then parallelly subjected to human miRNA array assay. In-depth data analysis from miRNA microarray data resulted in a list of 16 downregulated and 10 upregulated miRNAs whose transcripts are statistically significant with fold changes greater than 2 by TRPM7 knock-down. (B,C) Validation of miRNA array by siTRPM7 by qPCR. miR-26b-5p, miR-4530, and miR-28-5p were validated by real-time miRNA quantification using stem-loop RT-PCR in glioma cell lines A172 (B) and SNB19 (C) . A set of endogenous RNAs SNOD48, SNOD44, SNOD47, and U6 served as endogenous controls in real-time miRNA quantification to normalize miRNA gene expression. (D) Validation of miRNA array by overexpressing TRPM7. The constructs of wt hTRPM7 (M7-WT), Δ kinase (M7-DK), K1648R (MT-KR), and control (Ctrl) were introduced into and expressed in glioma cell lines A172, U373MG, SNB19, and U87MG for 48 h. Transfection efficiency was determined by TRPM7 mRNA expression detected by qRT-PCR. GAPDH was used as an internal control to monitor the presence of amplified cDNA in samples (upper left). Western blots by overexpression of individual TRPM7 constructs confirmed the data from qRT-PCR, which showed significantly increased TRPM7 in M7-WT, M7-KR, and M7-DK compared to the control ( D , right). The levels of miR-28-5p were determined by stem-loop RT-PCR (lower left). All the data were represented as fold changes relative to the appropriate controls and are the mean ± SD of triplicated samples performed in three independent experiments. (* P < 0.05, ** P < 0.01).
    Figure Legend Snippet: TRPM7 negatively regulates miR-28-5p in glioma cells. (A) Total RNA from biological triplicate samples, either with scramble control (siCtrl, A423, A424, and A425) or siTRPM7 (A427, A428, and A434), were extracted and then parallelly subjected to human miRNA array assay. In-depth data analysis from miRNA microarray data resulted in a list of 16 downregulated and 10 upregulated miRNAs whose transcripts are statistically significant with fold changes greater than 2 by TRPM7 knock-down. (B,C) Validation of miRNA array by siTRPM7 by qPCR. miR-26b-5p, miR-4530, and miR-28-5p were validated by real-time miRNA quantification using stem-loop RT-PCR in glioma cell lines A172 (B) and SNB19 (C) . A set of endogenous RNAs SNOD48, SNOD44, SNOD47, and U6 served as endogenous controls in real-time miRNA quantification to normalize miRNA gene expression. (D) Validation of miRNA array by overexpressing TRPM7. The constructs of wt hTRPM7 (M7-WT), Δ kinase (M7-DK), K1648R (MT-KR), and control (Ctrl) were introduced into and expressed in glioma cell lines A172, U373MG, SNB19, and U87MG for 48 h. Transfection efficiency was determined by TRPM7 mRNA expression detected by qRT-PCR. GAPDH was used as an internal control to monitor the presence of amplified cDNA in samples (upper left). Western blots by overexpression of individual TRPM7 constructs confirmed the data from qRT-PCR, which showed significantly increased TRPM7 in M7-WT, M7-KR, and M7-DK compared to the control ( D , right). The levels of miR-28-5p were determined by stem-loop RT-PCR (lower left). All the data were represented as fold changes relative to the appropriate controls and are the mean ± SD of triplicated samples performed in three independent experiments. (* P < 0.05, ** P < 0.01).

    Techniques Used: Control, Microarray, Knockdown, Biomarker Discovery, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Construct, Transfection, Expressing, Quantitative RT-PCR, Amplification, Western Blot, Over Expression

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    Article Snippet: .. The microRNA microarray was performed by LC Sciences (Houston, TX; RRID:SCR_000140) using μParafloTM chip technology with probes for all miRNA listed in Sanger miRBase Release 21 (mirBase.org, RRID:SCR_003152)). .. miR-146a-5p was quantified from RNA via reverse transcriptase qPCR, using miRCURY LNA miRNA SYBR Green RT-PCR Kit (Qiagen #339345) with miR-146a-5p-specific primers (Qiagen #YP00204688) as per the manufacturer’s instructions.

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    Isolation:

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    pcr-based mirna microarray platform  (Qiagen)
    90
    Qiagen pcr-based mirna microarray platform
    Total RNA was isolated from WT and IL10KO mice BM-FPCs using miRNeasy kit (Qiagen) and fibrosis-associated <t>miRNA</t> platform was used to assess the miRNAs level using Taqman microRNA assay. (A) At basal level fibrosis-associated miRNAs expression was significantly increased in IL10KO BM-FPCs compared to WT BM-FPCs with maximum expression of miRNA21, 145 and 208a (N=6). (B–D) miRNA21/145/208a expression was measured in WT-FPCs treated with IL10 and TGFβ by RT-PCR. IL10 treatment significantly inhibits TGFβ-induced miRNA-21, -145 and 208a expression. **p<0.01 vs BSA, #p<0.05 vs TGFβ. IL10KO mice: IL10 Knockout mice; BM-FPC: Bone marrow fibroblast progenitor cells. (N=6)
    Pcr Based Mirna Microarray Platform, supplied by Qiagen, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcr-based+mirna+microarray+platform/pcr+based+mirna+microarray+platform/pmc05736130-291-16-20
    Average 90 stars, based on 1 article reviews
    pcr-based mirna microarray platform - by Bioz Stars, 2026-11
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    Total RNA was isolated from WT and IL10KO mice BM-FPCs using miRNeasy kit (Qiagen) and fibrosis-associated miRNA platform was used to assess the miRNAs level using Taqman microRNA assay. (A) At basal level fibrosis-associated miRNAs expression was significantly increased in IL10KO BM-FPCs compared to WT BM-FPCs with maximum expression of miRNA21, 145 and 208a (N=6). (B–D) miRNA21/145/208a expression was measured in WT-FPCs treated with IL10 and TGFβ by RT-PCR. IL10 treatment significantly inhibits TGFβ-induced miRNA-21, -145 and 208a expression. **p<0.01 vs BSA, #p<0.05 vs TGFβ. IL10KO mice: IL10 Knockout mice; BM-FPC: Bone marrow fibroblast progenitor cells. (N=6)

    Journal: Circulation

    Article Title: Interleukin 10 Inhibits Bone Marrow Fibroblast Progenitor Cell-mediated Cardiac Fibrosis in Pressure Overloaded Myocardium

    doi: 10.1161/CIRCULATIONAHA.117.027889

    Figure Lengend Snippet: Total RNA was isolated from WT and IL10KO mice BM-FPCs using miRNeasy kit (Qiagen) and fibrosis-associated miRNA platform was used to assess the miRNAs level using Taqman microRNA assay. (A) At basal level fibrosis-associated miRNAs expression was significantly increased in IL10KO BM-FPCs compared to WT BM-FPCs with maximum expression of miRNA21, 145 and 208a (N=6). (B–D) miRNA21/145/208a expression was measured in WT-FPCs treated with IL10 and TGFβ by RT-PCR. IL10 treatment significantly inhibits TGFβ-induced miRNA-21, -145 and 208a expression. **p<0.01 vs BSA, #p<0.05 vs TGFβ. IL10KO mice: IL10 Knockout mice; BM-FPC: Bone marrow fibroblast progenitor cells. (N=6)

    Article Snippet: Fibrosis-associated miRNA expression was analyzed in WT and IL10 KO BM-FPCs at basal level using a PCR-based miRNA microarray platform (Qiagen). miRNA-array data showed that several miRNAs were differentially expressed in IL10KO BM-FPCs compared to WT BM-FPCs ( ).

    Techniques: Isolation, TaqMan microRNA Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Knock-Out

    BM-FPCs were transfected with miRNA-21 mimic for 12 hours using lipofectamine 2000 transfection reagent before IL10 and TGFβ treatments and miRNA-21 expression was measured by RT-PCR. (A) miRNA-21 mimic transfection significantly increased miRNA-21 expression in BM-FPCs. (B–E) 24 hrs after mimic, IL10 and TGFβ treatments, cells were harvested to measured the expression of fibrosis marker genes by RT-PCR. Relative levels of target genes expression were normalized with 18S. IL10 significantly inhibited TGFβ-induced fibrosis-associated genes expression. Restoration of miRNA-21 expression using miRNA-21 mimic significantly attenuated the IL10 effect on expression of these genes. ***p<0.001, **p<0.01, *p<0.05 vs BSA, ###p<0.001, ##p<0.01 vs TGFβ; $$$p<0.001, $p<0.05 vs TGFβ and ^^^p<0.001, ^^p<0.01, ^p<0.05 vs miR-21 mimic alone. (N=6) (F) Schematic diagram indicates TAC-induced mobilization and homing of BM-FPCs to the heart. In the heart, profibrotic stimuli (TGFβ in this case) first facilitates BM-FPC activation to myofibroblasts and then promote Smad2/3 dependent miRNA-21 maturation. The miRNA-21 maturation ultimately leads to fibrotic gene activation in activated myofibroblasts. On the other hand, IL10 inhibits TAC-induced mobilization, homing and activation of these cells and regulates cardiac fibrosis. Factor: Chemokine/cytokines; BMCs: Bone marrow cells; IL10 Interleukin-10.

    Journal: Circulation

    Article Title: Interleukin 10 Inhibits Bone Marrow Fibroblast Progenitor Cell-mediated Cardiac Fibrosis in Pressure Overloaded Myocardium

    doi: 10.1161/CIRCULATIONAHA.117.027889

    Figure Lengend Snippet: BM-FPCs were transfected with miRNA-21 mimic for 12 hours using lipofectamine 2000 transfection reagent before IL10 and TGFβ treatments and miRNA-21 expression was measured by RT-PCR. (A) miRNA-21 mimic transfection significantly increased miRNA-21 expression in BM-FPCs. (B–E) 24 hrs after mimic, IL10 and TGFβ treatments, cells were harvested to measured the expression of fibrosis marker genes by RT-PCR. Relative levels of target genes expression were normalized with 18S. IL10 significantly inhibited TGFβ-induced fibrosis-associated genes expression. Restoration of miRNA-21 expression using miRNA-21 mimic significantly attenuated the IL10 effect on expression of these genes. ***p<0.001, **p<0.01, *p<0.05 vs BSA, ###p<0.001, ##p<0.01 vs TGFβ; $$$p<0.001, $p<0.05 vs TGFβ and ^^^p<0.001, ^^p<0.01, ^p<0.05 vs miR-21 mimic alone. (N=6) (F) Schematic diagram indicates TAC-induced mobilization and homing of BM-FPCs to the heart. In the heart, profibrotic stimuli (TGFβ in this case) first facilitates BM-FPC activation to myofibroblasts and then promote Smad2/3 dependent miRNA-21 maturation. The miRNA-21 maturation ultimately leads to fibrotic gene activation in activated myofibroblasts. On the other hand, IL10 inhibits TAC-induced mobilization, homing and activation of these cells and regulates cardiac fibrosis. Factor: Chemokine/cytokines; BMCs: Bone marrow cells; IL10 Interleukin-10.

    Article Snippet: Fibrosis-associated miRNA expression was analyzed in WT and IL10 KO BM-FPCs at basal level using a PCR-based miRNA microarray platform (Qiagen). miRNA-array data showed that several miRNAs were differentially expressed in IL10KO BM-FPCs compared to WT BM-FPCs ( ).

    Techniques: Transfection, Expressing, Reverse Transcription Polymerase Chain Reaction, Marker, Activation Assay