rnase Search Results


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  • 99
    Thermo Fisher ribonuclease rnase
    ( A ) Representative sequencing gel exhibiting cleavage patterns of 32 P-labeled (*)hU2 and *hU6 snRNA paired with respective counter-strands subjected to enzymatic probing by ribonucleases <t>RNase</t> A and T1 (see Materials and Methods for experimental details).
    Ribonuclease Rnase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore rnase ribonuclease inhibitor
    ( A ) Representative sequencing gel exhibiting cleavage patterns of 32 P-labeled (*)hU2 and *hU6 snRNA paired with respective counter-strands subjected to enzymatic probing by ribonucleases <t>RNase</t> A and T1 (see Materials and Methods for experimental details).
    Rnase Ribonuclease Inhibitor, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Becton Dickinson ribonuclease rnase
    ( A ) Representative sequencing gel exhibiting cleavage patterns of 32 P-labeled (*)hU2 and *hU6 snRNA paired with respective counter-strands subjected to enzymatic probing by ribonucleases <t>RNase</t> A and T1 (see Materials and Methods for experimental details).
    Ribonuclease Rnase, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Interchim ribonuclease rnase
    ( A ) Representative sequencing gel exhibiting cleavage patterns of 32 P-labeled (*)hU2 and *hU6 snRNA paired with respective counter-strands subjected to enzymatic probing by ribonucleases <t>RNase</t> A and T1 (see Materials and Methods for experimental details).
    Ribonuclease Rnase, supplied by Interchim, used in various techniques. Bioz Stars score: 90/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Roche ribonuclease rnase
    ( A ) Representative sequencing gel exhibiting cleavage patterns of 32 P-labeled (*)hU2 and *hU6 snRNA paired with respective counter-strands subjected to enzymatic probing by ribonucleases <t>RNase</t> A and T1 (see Materials and Methods for experimental details).
    Ribonuclease Rnase, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore ribonuclease rnase
    (a) In vitro cell viability assay of B16F10 cancer cells incubated with PF-HQ, <t>DOX,</t> and PF-HQ–DOX in a dose-dependent manner for 24 h of treatment. The numerical value in the abscissa represents the concentration (μM) of DOX treatment. (b) Representative bright field images of B16F10 cells incubated with PF-HQ, DOX (2.5 μM), and PF-HQ–DOX (2.5 μM w.r.t. DOX) for 24 h were taken using an inverted microscope at 10× magnification. Scale bar = 50 microns. (c–g) The total DNA content was analyzed using <t>PI/RNase</t> by FACS in (c) untreated B16F10 cells as a control, and B16F10 cells treated with (d) free DOX (2.5 μM), (e) PF-HQ, or (f) PF-HQ–DOX (2.5 μM w.r.t. DOX) for 24 h. (g) Quantification of cell cycle analysis. (h–l) Analysis of apoptosis by flow cytometry using Annexin V-FITC assay in B16F10 cells treated with (i) DOX (2.5 μM), (j) PF-HQ, and (k) PF-HQ–DOX (2.5 μM w.r.t. DOX). (h) Untreated control cells were kept as a control. (l) Quantification data of apoptosis.
    Ribonuclease Rnase, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 81 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore rnase digestion
    NXF1 is present at the NPC independently of mRNA. (A) PolyA+ RNA detection by RNA FISH in U2OS cells. NXF1 (magenta) is detected in the NPCs while the mRNA (green) is not. The plot shows the intensity of both channels along the line in the merge. (B) PolyA+ RNA detection by RNA FISH in control cells and cells treated with <t>DRB</t> followed by <t>RNase</t> A digestion (RNase was added after permeabilization with 0.5% Triton, 2 min) and stained for NXF1. (C) Immunofluorescence of hnRNP A1, UAP56, CBP80, and NXF1 in control or DRB-treated U2OS cells. Decomposition of nucleoli is seen in the differential interference contrast (DIC) images indicating DRB transcription inhibition. (D) U2OS cell expressing the mutant Myc-NXF1-10RA protein that cannot bind mRNA (anti-Myc) and anti-Nup358 immunofluorescence for NPC detection. Cells were imaged at the middle (mid) or the top planes of the nucleus (top), and merged images show the presence of NXF1-10RA mutant in the NPCs. (E) NXF1 in control cells (top) or WGA-treated cells (bottom). Scale bars, 5 µm.
    Rnase Digestion, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 189 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Promega rnase one ribonuclease
    Acetylation of Ile-tRNA Ile , Val-tRNA Val and Met-tRNA Met isoacceptors in vivo . ( A ) Schematic diagram of detection and quantification of <t>acetyl-aminoacyl-tRNAs,</t> produced by the action of ItaT in vivo (See details in Materials and Methods). ( B ) LC/MS analysis of <t>RNase</t> I-digested fragments of acetyl-aminoacyl-tRNAs. Identification of the molecular masses corresponding to Ac-Ile/Leu-A76 (acetyl-isoleucyl/leucyl-adenosine, m/z 423.19), Ac-Val-A76 (acetyl-valyl-adenosine, m / z 409.18) and Ac-Met-A76(acetyl-methionyl-adenosine, m / z 441.15) derived from Ac-Ile/Leu-tRNA Ile/Leu , Ac-Val-tRNA Val and Ac-Met-tRNA Met , respectively, produced by the action of ItaT in vivo . Ac-Ala-A76 (acetyl-alanyl-adenosine) and Ac-Phe-A76 (acetyl-phenylalanyl-adenosine) were not detected. Identification of the molecular masses corresponding to D 3 Ac-Ile/Leu-A76 ( m/z 426.21), D 3 Ac-Val-A76 ( m/z 412.20), D 3 Ac-Met-A76 ( m/z 444.17), D 3 Ac-Ala-A76 ( m/z 384.17) and D 3 Ac-Phe-A76 ( m/z 460.20), derived from the in vitro aminoacyl-tRNAs chemically acetylated by acetic anhydride-D 6 . The two observed peaks in each Ac-aa-A76 represent structural isomers of 3′-acetyl-aminoacyl-A76 and 2′-acetyl-aminoacyl-A76, as observed for the separation of 3′-O-methyl and 2′-O-methyl nucleosides ( 38 ). ( C ) Quantification of the acetylation of aminoacyl-tRNAs by the action of ItaT in vivo . The fractions of individual acetylated aminoacyl-tRNAs by ItaT in vivo were estimated as the ratio of the amount of Ac-aa-A76 to the sum of the amounts of Ac-aa-A76 and D 3 Ac-aa-A76. The bars in the graphs are SD of more than three independent experiments.
    Rnase One Ribonuclease, supplied by Promega, used in various techniques. Bioz Stars score: 88/100, based on 106 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Promega rnase ribonuclease
    Acetylation of Ile-tRNA Ile , Val-tRNA Val and Met-tRNA Met isoacceptors in vivo . ( A ) Schematic diagram of detection and quantification of <t>acetyl-aminoacyl-tRNAs,</t> produced by the action of ItaT in vivo (See details in Materials and Methods). ( B ) LC/MS analysis of <t>RNase</t> I-digested fragments of acetyl-aminoacyl-tRNAs. Identification of the molecular masses corresponding to Ac-Ile/Leu-A76 (acetyl-isoleucyl/leucyl-adenosine, m/z 423.19), Ac-Val-A76 (acetyl-valyl-adenosine, m / z 409.18) and Ac-Met-A76(acetyl-methionyl-adenosine, m / z 441.15) derived from Ac-Ile/Leu-tRNA Ile/Leu , Ac-Val-tRNA Val and Ac-Met-tRNA Met , respectively, produced by the action of ItaT in vivo . Ac-Ala-A76 (acetyl-alanyl-adenosine) and Ac-Phe-A76 (acetyl-phenylalanyl-adenosine) were not detected. Identification of the molecular masses corresponding to D 3 Ac-Ile/Leu-A76 ( m/z 426.21), D 3 Ac-Val-A76 ( m/z 412.20), D 3 Ac-Met-A76 ( m/z 444.17), D 3 Ac-Ala-A76 ( m/z 384.17) and D 3 Ac-Phe-A76 ( m/z 460.20), derived from the in vitro aminoacyl-tRNAs chemically acetylated by acetic anhydride-D 6 . The two observed peaks in each Ac-aa-A76 represent structural isomers of 3′-acetyl-aminoacyl-A76 and 2′-acetyl-aminoacyl-A76, as observed for the separation of 3′-O-methyl and 2′-O-methyl nucleosides ( 38 ). ( C ) Quantification of the acetylation of aminoacyl-tRNAs by the action of ItaT in vivo . The fractions of individual acetylated aminoacyl-tRNAs by ItaT in vivo were estimated as the ratio of the amount of Ac-aa-A76 to the sum of the amounts of Ac-aa-A76 and D 3 Ac-aa-A76. The bars in the graphs are SD of more than three independent experiments.
    Rnase Ribonuclease, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore ribonuclease rnase a
     Cellular RNA is required for efficient interaction between HuD molecules. ( A ) Scheme of the  in vitro  binding experiment using HeLa cell extracts containing T7–HuD and FLAG–HuD. ( B ) Immunoprecipitation of the mixed extracts with anti-T7 antibody in the presence or absence of RNase A. RNase treatment had no effect on the levels of tagged HuD in the extract (two bottom panels). The asterisks indicate the heavy and light chains of IgG used for immunoprecipitation. ( C ) Binding of wild-type HuD and a mutant HuDmt to the poly(U) sequence. HeLa cell extracts containing T7–HuD or T7–HuDmt were incubated with poly(U)–Sepharose beads and the bound proteins were analyzed by western blotting with anti-T7 antibody. As controls, the levels of T7–HuD and T7–HuDmt in the extracts used are shown (input). ( D ) Pull-down of the  in vitro  translated myc-HuD or myc-HuDmt by GST or GST–HuD. Proteins pulled-down by GST or GST–HuD were analyzed by western blotting with anti-myc antibody. As controls, the levels of myc-HuD and myc-HuDmt in the  in vitro  translation reactions used are shown (input).
    Ribonuclease Rnase A, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 56 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Promega ribonuclease rnase a
     Cellular RNA is required for efficient interaction between HuD molecules. ( A ) Scheme of the  in vitro  binding experiment using HeLa cell extracts containing T7–HuD and FLAG–HuD. ( B ) Immunoprecipitation of the mixed extracts with anti-T7 antibody in the presence or absence of RNase A. RNase treatment had no effect on the levels of tagged HuD in the extract (two bottom panels). The asterisks indicate the heavy and light chains of IgG used for immunoprecipitation. ( C ) Binding of wild-type HuD and a mutant HuDmt to the poly(U) sequence. HeLa cell extracts containing T7–HuD or T7–HuDmt were incubated with poly(U)–Sepharose beads and the bound proteins were analyzed by western blotting with anti-T7 antibody. As controls, the levels of T7–HuD and T7–HuDmt in the extracts used are shown (input). ( D ) Pull-down of the  in vitro  translated myc-HuD or myc-HuDmt by GST or GST–HuD. Proteins pulled-down by GST or GST–HuD were analyzed by western blotting with anti-myc antibody. As controls, the levels of myc-HuD and myc-HuDmt in the  in vitro  translation reactions used are shown (input).
    Ribonuclease Rnase A, supplied by Promega, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Qiagen rnase
    Characterization of the nature and the source of the RIBE factors a, b, Treatment of UV-CM and UV-Ctrl collected from N2 animals irradiated at 100 J/m 2 with <t>RNase</t> (1μg/μL) or <t>DNase</t> (0.01 Unit/μL) did not alter the apoptosis-inhibitory effect on ced-1(e1735) animals (Methods). Germ cell corpses were scored after 48-hour treatment of ced-1(e1735) L4 larvae. c, e, f, g, ced-1(e1735) L4 larvae were treated with UV-CM and UV-control (0.1 μg/μL) prepared from ced-3(n2433) animals ( c ), glp-1(e2141ts) animals grown at 25°C ( e ), N2 animals fed with formaldehyde-treated HB101 bacteria ( f ), and P cpr-4::cpr-4::flag; cpr-4(tm3718) animals with or without anti-Flag depletion ( g ), respectively. Data are mean ± s.e.m. The numbers of gonad arms scored are indicated inside the bars ( a–c, e–g ). ** P
    Rnase, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 3800 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Promega rnase onetm ribonuclease
    Characterization of the nature and the source of the RIBE factors a, b, Treatment of UV-CM and UV-Ctrl collected from N2 animals irradiated at 100 J/m 2 with <t>RNase</t> (1μg/μL) or <t>DNase</t> (0.01 Unit/μL) did not alter the apoptosis-inhibitory effect on ced-1(e1735) animals (Methods). Germ cell corpses were scored after 48-hour treatment of ced-1(e1735) L4 larvae. c, e, f, g, ced-1(e1735) L4 larvae were treated with UV-CM and UV-control (0.1 μg/μL) prepared from ced-3(n2433) animals ( c ), glp-1(e2141ts) animals grown at 25°C ( e ), N2 animals fed with formaldehyde-treated HB101 bacteria ( f ), and P cpr-4::cpr-4::flag; cpr-4(tm3718) animals with or without anti-Flag depletion ( g ), respectively. Data are mean ± s.e.m. The numbers of gonad arms scored are indicated inside the bars ( a–c, e–g ). ** P
    Rnase Onetm Ribonuclease, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Millipore rnase inhibitor
    m 6 A alters RNA structure to recruit <t>HNRNPG.</t> ( A ) Sequence logo of the most enriched motif within HNRNPG PAR-CLIP peaks. ( B ) Left: secondary structure of the MALAT1 hairpin, showing the A-2,515-to-G/C/U mutations that were introduced at the m 6 A site. Right: quantification of relative HNRNPG pull-down with the original (2,515-A) and mutated (2,515-G/C/U) MALAT1 hairpins, normalized to pulled-down HIST1H1C. Data shown as mean; error bar = standard deviation; n = 3 biological replicates. ( C ) Left: structural probing of the unmethylated and methylated MALAT1 hairpins. The orange lines indicate regions with marked differences between the unmethylated and methylated hairpins. The location of the m 6 A residue is indicated by a red dot. Ctrl, no nuclease added; V1; <t>RNase</t> V1 digestion; S1, S1 nuclease digestion; T1, RNase T1 digestion; G-L, G-ladder; AH, alkaline hydrolysis. Right: secondary structure of the unmethylated and methylated MALAT1 hairpins, marked at their S1 nuclease (red lines) and V1 nuclease (green lines) cleavage sites. ( D ) Model showing that m 6 A disrupts RNA structure, exposes a motif that includes the m 6 A site, and recruits an RNA binding protein.
    Rnase Inhibitor, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1112 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher rnase
    In vitro cell proliferation and anti-apoptotic effects of <t>MPs.</t> (A) Proliferation of HUVEC treated with vehicle control, with different doses of MPs or MP-treated with <t>RNase</t> (MP-RNase). (B) Apoptosis assay of HUVEC treated with vehicle control or with different doses of MPs. (C) Apoptosis assay of HMVEC treated with vehicle control or with different doses of MPs. Results are expressed as % compared to vehicle control and mean ± SE of three different experiments. Kruskal-Wallis test was performed; * P
    Rnase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 9347 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    ( A ) Representative sequencing gel exhibiting cleavage patterns of 32 P-labeled (*)hU2 and *hU6 snRNA paired with respective counter-strands subjected to enzymatic probing by ribonucleases RNase A and T1 (see Materials and Methods for experimental details).

    Journal: RNA

    Article Title: Conformational heterogeneity of the protein-free human spliceosomal U2-U6 snRNA complex

    doi: 10.1261/rna.038265.113

    Figure Lengend Snippet: ( A ) Representative sequencing gel exhibiting cleavage patterns of 32 P-labeled (*)hU2 and *hU6 snRNA paired with respective counter-strands subjected to enzymatic probing by ribonucleases RNase A and T1 (see Materials and Methods for experimental details).

    Article Snippet: The resulting complex was subjected to partial cleavage by ribonucleases RNase A, T1 (both from Ambion, Inc.), and RNase V1 (Pierce Milwaukee).

    Techniques: Sequencing, Labeling

    (a) In vitro cell viability assay of B16F10 cancer cells incubated with PF-HQ, DOX, and PF-HQ–DOX in a dose-dependent manner for 24 h of treatment. The numerical value in the abscissa represents the concentration (μM) of DOX treatment. (b) Representative bright field images of B16F10 cells incubated with PF-HQ, DOX (2.5 μM), and PF-HQ–DOX (2.5 μM w.r.t. DOX) for 24 h were taken using an inverted microscope at 10× magnification. Scale bar = 50 microns. (c–g) The total DNA content was analyzed using PI/RNase by FACS in (c) untreated B16F10 cells as a control, and B16F10 cells treated with (d) free DOX (2.5 μM), (e) PF-HQ, or (f) PF-HQ–DOX (2.5 μM w.r.t. DOX) for 24 h. (g) Quantification of cell cycle analysis. (h–l) Analysis of apoptosis by flow cytometry using Annexin V-FITC assay in B16F10 cells treated with (i) DOX (2.5 μM), (j) PF-HQ, and (k) PF-HQ–DOX (2.5 μM w.r.t. DOX). (h) Untreated control cells were kept as a control. (l) Quantification data of apoptosis.

    Journal: Chemical Science

    Article Title: Multifunctional (3-in-1) cancer theranostics applications of hydroxyquinoline-appended polyfluorene nanoparticles (3-in-1) cancer theranostics applications of hydroxyquinoline-appended polyfluorene nanoparticles †Electronic supplementary information (ESI) available: Synthesis, characterization, stability, optical properties, imaging, drug delivery, etc. See DOI: 10.1039/c7sc03321d

    doi: 10.1039/c7sc03321d

    Figure Lengend Snippet: (a) In vitro cell viability assay of B16F10 cancer cells incubated with PF-HQ, DOX, and PF-HQ–DOX in a dose-dependent manner for 24 h of treatment. The numerical value in the abscissa represents the concentration (μM) of DOX treatment. (b) Representative bright field images of B16F10 cells incubated with PF-HQ, DOX (2.5 μM), and PF-HQ–DOX (2.5 μM w.r.t. DOX) for 24 h were taken using an inverted microscope at 10× magnification. Scale bar = 50 microns. (c–g) The total DNA content was analyzed using PI/RNase by FACS in (c) untreated B16F10 cells as a control, and B16F10 cells treated with (d) free DOX (2.5 μM), (e) PF-HQ, or (f) PF-HQ–DOX (2.5 μM w.r.t. DOX) for 24 h. (g) Quantification of cell cycle analysis. (h–l) Analysis of apoptosis by flow cytometry using Annexin V-FITC assay in B16F10 cells treated with (i) DOX (2.5 μM), (j) PF-HQ, and (k) PF-HQ–DOX (2.5 μM w.r.t. DOX). (h) Untreated control cells were kept as a control. (l) Quantification data of apoptosis.

    Article Snippet: Materials Doxorubicin (DOX), Dulbecco’s Modified Eagle’s medium (DMEM), phosphate-buffered saline (PBS), kanamycin, streptomycin, penicillin, ribonuclease (RNase), fetal bovine serum (FBS), HBSS buffer (Hank’s balanced salt solution), MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), and propidium iodide (PI) were procured from Sigma Aldrich Chemicals, USA and used without any purification.

    Techniques: In Vitro, Viability Assay, Incubation, Concentration Assay, Inverted Microscopy, FACS, Cell Cycle Assay, Flow Cytometry, Cytometry

    Translation events in Tau-positive neurites are a result of local protein synthesis (A) Cells grown for 9 DIV and treated with DMSO for 24 h. Cells immunostained with an anti-Tau antibody (magenta) were incubated with SYTO RNASelect green fluorescent dye to label endogenous RNA (green). Total green fluorescence intensity was measured in neurites covering a distance of 150 μm from the edge of the soma (2, + SYTO). As negative control, green fluorescence was measured in cells that had not been incubated with SYTO (1, -SYTO). To determine if SYTO selectively labeled RNA, some fixed cells were digested with DNAse (3, +SYTO +DNAse) or with RNAse (4, +SYTO +RNAse). Box and whisker graphs represent the average relative fluorescence intensity of 10 neurites per condition, shown as individual data points, and the mean and median of 5 (n=5, -SYTO negative samples compared to their corresponding +SYTO controls) or 6 (n=6, +SYTO + DNAse and +SYTO +RNAse compared to their corresponding + SYTO controls) independent experiments. *** p

    Journal: bioRxiv

    Article Title: Object-based analyses in FIJI/ImageJ to measure local RNA translation sites in neurites in response to Aβ1-42 oligomers

    doi: 10.1101/2020.01.27.921494

    Figure Lengend Snippet: Translation events in Tau-positive neurites are a result of local protein synthesis (A) Cells grown for 9 DIV and treated with DMSO for 24 h. Cells immunostained with an anti-Tau antibody (magenta) were incubated with SYTO RNASelect green fluorescent dye to label endogenous RNA (green). Total green fluorescence intensity was measured in neurites covering a distance of 150 μm from the edge of the soma (2, + SYTO). As negative control, green fluorescence was measured in cells that had not been incubated with SYTO (1, -SYTO). To determine if SYTO selectively labeled RNA, some fixed cells were digested with DNAse (3, +SYTO +DNAse) or with RNAse (4, +SYTO +RNAse). Box and whisker graphs represent the average relative fluorescence intensity of 10 neurites per condition, shown as individual data points, and the mean and median of 5 (n=5, -SYTO negative samples compared to their corresponding +SYTO controls) or 6 (n=6, +SYTO + DNAse and +SYTO +RNAse compared to their corresponding + SYTO controls) independent experiments. *** p

    Article Snippet: Some fixed neurons were incubated with 50 μg/ml DNAse or RNAse (Sigma) for 10 minutes at room temperature to assess the selectivity of the SYTO labeling.

    Techniques: Incubation, Fluorescence, Negative Control, Labeling, Whisker Assay

    NXF1 is present at the NPC independently of mRNA. (A) PolyA+ RNA detection by RNA FISH in U2OS cells. NXF1 (magenta) is detected in the NPCs while the mRNA (green) is not. The plot shows the intensity of both channels along the line in the merge. (B) PolyA+ RNA detection by RNA FISH in control cells and cells treated with DRB followed by RNase A digestion (RNase was added after permeabilization with 0.5% Triton, 2 min) and stained for NXF1. (C) Immunofluorescence of hnRNP A1, UAP56, CBP80, and NXF1 in control or DRB-treated U2OS cells. Decomposition of nucleoli is seen in the differential interference contrast (DIC) images indicating DRB transcription inhibition. (D) U2OS cell expressing the mutant Myc-NXF1-10RA protein that cannot bind mRNA (anti-Myc) and anti-Nup358 immunofluorescence for NPC detection. Cells were imaged at the middle (mid) or the top planes of the nucleus (top), and merged images show the presence of NXF1-10RA mutant in the NPCs. (E) NXF1 in control cells (top) or WGA-treated cells (bottom). Scale bars, 5 µm.

    Journal: The Journal of Cell Biology

    Article Title: Imaging within single NPCs reveals NXF1’s role in mRNA export on the cytoplasmic side of the pore

    doi: 10.1083/jcb.201901127

    Figure Lengend Snippet: NXF1 is present at the NPC independently of mRNA. (A) PolyA+ RNA detection by RNA FISH in U2OS cells. NXF1 (magenta) is detected in the NPCs while the mRNA (green) is not. The plot shows the intensity of both channels along the line in the merge. (B) PolyA+ RNA detection by RNA FISH in control cells and cells treated with DRB followed by RNase A digestion (RNase was added after permeabilization with 0.5% Triton, 2 min) and stained for NXF1. (C) Immunofluorescence of hnRNP A1, UAP56, CBP80, and NXF1 in control or DRB-treated U2OS cells. Decomposition of nucleoli is seen in the differential interference contrast (DIC) images indicating DRB transcription inhibition. (D) U2OS cell expressing the mutant Myc-NXF1-10RA protein that cannot bind mRNA (anti-Myc) and anti-Nup358 immunofluorescence for NPC detection. Cells were imaged at the middle (mid) or the top planes of the nucleus (top), and merged images show the presence of NXF1-10RA mutant in the NPCs. (E) NXF1 in control cells (top) or WGA-treated cells (bottom). Scale bars, 5 µm.

    Article Snippet: For transcription inhibition, cells were treated with 100 µM DRB (Sigma) for 3 h. For RNase digestion, cells were treated with 5 µg/ml actinomycin D (Sigma) for 3 h and then permeabilized with 0.5% Triton X-100 in PBS for 2 min and digested with RNase A (100 µg/ml in PBS with 3 mM MgCl2 ; Sigma) for 45 min at room temperature.

    Techniques: RNA Detection, Fluorescence In Situ Hybridization, Staining, Immunofluorescence, Inhibition, Expressing, Mutagenesis, Whole Genome Amplification

    Fig. 3. CBTF 122 and CBTF 98 bind RNA in vitro and are associated with non-translating mRNAs in cleavage stage embryos. ( A ) Northwestern blot analysis of total soluble proteins from two stage VI oocytes (VI) or two embryos at the stages shown. Proteins were separated by SDS–PAGE and transferred to nitrocellulose. The blot was probed first with 32 P-labelled L1 5′-UTR (northwestern) and subsequently probed with affinity-purified anti-CBTF 122/98 antibody (western). ( B ) Cytoplasmic proteins from cleavage-stage embryos were incubated in the absence or presence of RNase A and T1 as indicated and then sedimented through a 20–60% nycodenz gradient. Protein precipitated from the indicated fractions was analysed by western blotting using affinity-purified anti-CBTF 122/98 antibody or anti-mRNP3+4 antibody. ( C ) Fractions 12 and 13 of the (–)RNase gradient were mixed and aliquots were incubated in the absence or presence of RNase A and T1. The RNase-treated or untreated aliquots were incubated with anti-mRNP3+4 antiserum (α-mRNP3+4) and protein A beads as shown. Western blots of the supernatant (S) and pellet (P) fractions were probed with affinity-purified anti-p122/p98 (upper panel) or anti-mRNP3+4 (lower panel) antibodies.

    Journal: The EMBO Journal

    Article Title: RNA-dependent cytoplasmic anchoring of a transcription factor subunit during Xenopus development

    doi: 10.1093/emboj/19.14.3683

    Figure Lengend Snippet: Fig. 3. CBTF 122 and CBTF 98 bind RNA in vitro and are associated with non-translating mRNAs in cleavage stage embryos. ( A ) Northwestern blot analysis of total soluble proteins from two stage VI oocytes (VI) or two embryos at the stages shown. Proteins were separated by SDS–PAGE and transferred to nitrocellulose. The blot was probed first with 32 P-labelled L1 5′-UTR (northwestern) and subsequently probed with affinity-purified anti-CBTF 122/98 antibody (western). ( B ) Cytoplasmic proteins from cleavage-stage embryos were incubated in the absence or presence of RNase A and T1 as indicated and then sedimented through a 20–60% nycodenz gradient. Protein precipitated from the indicated fractions was analysed by western blotting using affinity-purified anti-CBTF 122/98 antibody or anti-mRNP3+4 antibody. ( C ) Fractions 12 and 13 of the (–)RNase gradient were mixed and aliquots were incubated in the absence or presence of RNase A and T1. The RNase-treated or untreated aliquots were incubated with anti-mRNP3+4 antiserum (α-mRNP3+4) and protein A beads as shown. Western blots of the supernatant (S) and pellet (P) fractions were probed with affinity-purified anti-p122/p98 (upper panel) or anti-mRNP3+4 (lower panel) antibodies.

    Article Snippet: For RNase treatment, nuclear and embryonic extracts, and nycodenz gradient fractions were incubated in the presence of 100 µg/ml RNase A (Sigma) and 1500 U/ml RNase T1 (Amersham) at 37°C for 30 min either before centrifugation or before immunoprecipitation.

    Techniques: In Vitro, SDS Page, Affinity Purification, Western Blot, Incubation

    RNase and DNase activities of the recombinant Gly m 4l protein. (A) RNase activity of Gly m 4l protein under different pH conditions. Samples containing 10 μg total soybean RNA were incubated with the presence of 10 μg Gly m 4l protein at 37°C for 0, 2 and 4 h at pH 3, 5, 7 and 9 from left to right. (B) RNase activity of the recombinant Gly m 4l protein analyzed in the presence of zeatin using total soybean RNA at pH 7. Samples containing 10 μg total soybean RNA were incubated with the presence of 10 μg Gly m 4l protein and 10 mM zeatin at 37°C for 30 min, 1, 2, 3 and 4 h. The samples incubated with the presence of the recombinant Gly m 4l protein or boiled Gly m 4l protein and zeatin were used as negative controls. (C) RNase activity of the recombinant Gly m 4l protein analyzed in the presence of zeatin using yeast tRNA. Samples containing 100 μg yeast tRNA were incubated with the presence of 10 μg recombinant Gly m 4l protein and 10 mM zeatin at 37°C for 30 min. The samples incubated with the presence of Gly m 4l protein alone or boiled Gly m 4l protein and zeatin were used as controls. The results were calculated by the 2 −ΔΔCt method. (D) DNase activity of the recombinant Gly m 4l protein under different pH conditions. Samples containing 10 μg total soybean DNA and 10 μg Gly m 4l protein were incubated at 37°C for 0 and 4 h at pH 3, 5, 7 and 9 from left to right. The experiments were performed on three technical replicates and statistically analysed using Student’s t-test (*P

    Journal: PLoS ONE

    Article Title: A Novel Pathogenesis-Related Class 10 Protein Gly m 4l, Increases Resistance upon Phytophthora sojae Infection in Soybean (Glycine max [L.] Merr.)

    doi: 10.1371/journal.pone.0140364

    Figure Lengend Snippet: RNase and DNase activities of the recombinant Gly m 4l protein. (A) RNase activity of Gly m 4l protein under different pH conditions. Samples containing 10 μg total soybean RNA were incubated with the presence of 10 μg Gly m 4l protein at 37°C for 0, 2 and 4 h at pH 3, 5, 7 and 9 from left to right. (B) RNase activity of the recombinant Gly m 4l protein analyzed in the presence of zeatin using total soybean RNA at pH 7. Samples containing 10 μg total soybean RNA were incubated with the presence of 10 μg Gly m 4l protein and 10 mM zeatin at 37°C for 30 min, 1, 2, 3 and 4 h. The samples incubated with the presence of the recombinant Gly m 4l protein or boiled Gly m 4l protein and zeatin were used as negative controls. (C) RNase activity of the recombinant Gly m 4l protein analyzed in the presence of zeatin using yeast tRNA. Samples containing 100 μg yeast tRNA were incubated with the presence of 10 μg recombinant Gly m 4l protein and 10 mM zeatin at 37°C for 30 min. The samples incubated with the presence of Gly m 4l protein alone or boiled Gly m 4l protein and zeatin were used as controls. The results were calculated by the 2 −ΔΔCt method. (D) DNase activity of the recombinant Gly m 4l protein under different pH conditions. Samples containing 10 μg total soybean DNA and 10 μg Gly m 4l protein were incubated at 37°C for 0 and 4 h at pH 3, 5, 7 and 9 from left to right. The experiments were performed on three technical replicates and statistically analysed using Student’s t-test (*P

    Article Snippet: Meanwhile, the RNase activity of the recombinant Gly m 4l protein on yeast tRNA (Sigma, USA) was also assayed by determining the generation of acid-soluble, UV-absorbing species as described by Wang and Ng [ ].

    Techniques: Recombinant, Activity Assay, Incubation

    Acetylation of Ile-tRNA Ile , Val-tRNA Val and Met-tRNA Met isoacceptors in vivo . ( A ) Schematic diagram of detection and quantification of acetyl-aminoacyl-tRNAs, produced by the action of ItaT in vivo (See details in Materials and Methods). ( B ) LC/MS analysis of RNase I-digested fragments of acetyl-aminoacyl-tRNAs. Identification of the molecular masses corresponding to Ac-Ile/Leu-A76 (acetyl-isoleucyl/leucyl-adenosine, m/z 423.19), Ac-Val-A76 (acetyl-valyl-adenosine, m / z 409.18) and Ac-Met-A76(acetyl-methionyl-adenosine, m / z 441.15) derived from Ac-Ile/Leu-tRNA Ile/Leu , Ac-Val-tRNA Val and Ac-Met-tRNA Met , respectively, produced by the action of ItaT in vivo . Ac-Ala-A76 (acetyl-alanyl-adenosine) and Ac-Phe-A76 (acetyl-phenylalanyl-adenosine) were not detected. Identification of the molecular masses corresponding to D 3 Ac-Ile/Leu-A76 ( m/z 426.21), D 3 Ac-Val-A76 ( m/z 412.20), D 3 Ac-Met-A76 ( m/z 444.17), D 3 Ac-Ala-A76 ( m/z 384.17) and D 3 Ac-Phe-A76 ( m/z 460.20), derived from the in vitro aminoacyl-tRNAs chemically acetylated by acetic anhydride-D 6 . The two observed peaks in each Ac-aa-A76 represent structural isomers of 3′-acetyl-aminoacyl-A76 and 2′-acetyl-aminoacyl-A76, as observed for the separation of 3′-O-methyl and 2′-O-methyl nucleosides ( 38 ). ( C ) Quantification of the acetylation of aminoacyl-tRNAs by the action of ItaT in vivo . The fractions of individual acetylated aminoacyl-tRNAs by ItaT in vivo were estimated as the ratio of the amount of Ac-aa-A76 to the sum of the amounts of Ac-aa-A76 and D 3 Ac-aa-A76. The bars in the graphs are SD of more than three independent experiments.

    Journal: Nucleic Acids Research

    Article Title: Substrate specificities of Escherichia coli ItaT that acetylates aminoacyl-tRNAs

    doi: 10.1093/nar/gkaa487

    Figure Lengend Snippet: Acetylation of Ile-tRNA Ile , Val-tRNA Val and Met-tRNA Met isoacceptors in vivo . ( A ) Schematic diagram of detection and quantification of acetyl-aminoacyl-tRNAs, produced by the action of ItaT in vivo (See details in Materials and Methods). ( B ) LC/MS analysis of RNase I-digested fragments of acetyl-aminoacyl-tRNAs. Identification of the molecular masses corresponding to Ac-Ile/Leu-A76 (acetyl-isoleucyl/leucyl-adenosine, m/z 423.19), Ac-Val-A76 (acetyl-valyl-adenosine, m / z 409.18) and Ac-Met-A76(acetyl-methionyl-adenosine, m / z 441.15) derived from Ac-Ile/Leu-tRNA Ile/Leu , Ac-Val-tRNA Val and Ac-Met-tRNA Met , respectively, produced by the action of ItaT in vivo . Ac-Ala-A76 (acetyl-alanyl-adenosine) and Ac-Phe-A76 (acetyl-phenylalanyl-adenosine) were not detected. Identification of the molecular masses corresponding to D 3 Ac-Ile/Leu-A76 ( m/z 426.21), D 3 Ac-Val-A76 ( m/z 412.20), D 3 Ac-Met-A76 ( m/z 444.17), D 3 Ac-Ala-A76 ( m/z 384.17) and D 3 Ac-Phe-A76 ( m/z 460.20), derived from the in vitro aminoacyl-tRNAs chemically acetylated by acetic anhydride-D 6 . The two observed peaks in each Ac-aa-A76 represent structural isomers of 3′-acetyl-aminoacyl-A76 and 2′-acetyl-aminoacyl-A76, as observed for the separation of 3′-O-methyl and 2′-O-methyl nucleosides ( 38 ). ( C ) Quantification of the acetylation of aminoacyl-tRNAs by the action of ItaT in vivo . The fractions of individual acetylated aminoacyl-tRNAs by ItaT in vivo were estimated as the ratio of the amount of Ac-aa-A76 to the sum of the amounts of Ac-aa-A76 and D 3 Ac-aa-A76. The bars in the graphs are SD of more than three independent experiments.

    Article Snippet: LC/MS spectrometry The purified acetylated aminoacyl-tRNAs described above were digested with RNase One Ribonuclease (Promega, Japan), in a reaction mixture (25 μl volume) containing 25 mM NH4 OAc and 2.5 units enzyme, at 37°C for 60 min.

    Techniques: In Vivo, Produced, Liquid Chromatography with Mass Spectroscopy, Derivative Assay, In Vitro

     Cellular RNA is required for efficient interaction between HuD molecules. ( A ) Scheme of the  in vitro  binding experiment using HeLa cell extracts containing T7–HuD and FLAG–HuD. ( B ) Immunoprecipitation of the mixed extracts with anti-T7 antibody in the presence or absence of RNase A. RNase treatment had no effect on the levels of tagged HuD in the extract (two bottom panels). The asterisks indicate the heavy and light chains of IgG used for immunoprecipitation. ( C ) Binding of wild-type HuD and a mutant HuDmt to the poly(U) sequence. HeLa cell extracts containing T7–HuD or T7–HuDmt were incubated with poly(U)–Sepharose beads and the bound proteins were analyzed by western blotting with anti-T7 antibody. As controls, the levels of T7–HuD and T7–HuDmt in the extracts used are shown (input). ( D ) Pull-down of the  in vitro  translated myc-HuD or myc-HuDmt by GST or GST–HuD. Proteins pulled-down by GST or GST–HuD were analyzed by western blotting with anti-myc antibody. As controls, the levels of myc-HuD and myc-HuDmt in the  in vitro  translation reactions used are shown (input).

    Journal: Nucleic Acids Research

    Article Title: Complex formation of the neuron-specific ELAV-like Hu RNA-binding proteins

    doi:

    Figure Lengend Snippet: Cellular RNA is required for efficient interaction between HuD molecules. ( A ) Scheme of the in vitro binding experiment using HeLa cell extracts containing T7–HuD and FLAG–HuD. ( B ) Immunoprecipitation of the mixed extracts with anti-T7 antibody in the presence or absence of RNase A. RNase treatment had no effect on the levels of tagged HuD in the extract (two bottom panels). The asterisks indicate the heavy and light chains of IgG used for immunoprecipitation. ( C ) Binding of wild-type HuD and a mutant HuDmt to the poly(U) sequence. HeLa cell extracts containing T7–HuD or T7–HuDmt were incubated with poly(U)–Sepharose beads and the bound proteins were analyzed by western blotting with anti-T7 antibody. As controls, the levels of T7–HuD and T7–HuDmt in the extracts used are shown (input). ( D ) Pull-down of the in vitro translated myc-HuD or myc-HuDmt by GST or GST–HuD. Proteins pulled-down by GST or GST–HuD were analyzed by western blotting with anti-myc antibody. As controls, the levels of myc-HuD and myc-HuDmt in the in vitro translation reactions used are shown (input).

    Article Snippet: These extracts were separately incubated with or without ribonuclease (RNase) A (0.4 mg/ml; Sigma) for 1 h at 37°C.

    Techniques: In Vitro, Binding Assay, Immunoprecipitation, Mutagenesis, Sequencing, Incubation, Western Blot

    Characterization of the nature and the source of the RIBE factors a, b, Treatment of UV-CM and UV-Ctrl collected from N2 animals irradiated at 100 J/m 2 with RNase (1μg/μL) or DNase (0.01 Unit/μL) did not alter the apoptosis-inhibitory effect on ced-1(e1735) animals (Methods). Germ cell corpses were scored after 48-hour treatment of ced-1(e1735) L4 larvae. c, e, f, g, ced-1(e1735) L4 larvae were treated with UV-CM and UV-control (0.1 μg/μL) prepared from ced-3(n2433) animals ( c ), glp-1(e2141ts) animals grown at 25°C ( e ), N2 animals fed with formaldehyde-treated HB101 bacteria ( f ), and P cpr-4::cpr-4::flag; cpr-4(tm3718) animals with or without anti-Flag depletion ( g ), respectively. Data are mean ± s.e.m. The numbers of gonad arms scored are indicated inside the bars ( a–c, e–g ). ** P

    Journal: Nature

    Article Title: Cysteine protease cathepsin B mediates radiation-induced bystander effects

    doi: 10.1038/nature23284

    Figure Lengend Snippet: Characterization of the nature and the source of the RIBE factors a, b, Treatment of UV-CM and UV-Ctrl collected from N2 animals irradiated at 100 J/m 2 with RNase (1μg/μL) or DNase (0.01 Unit/μL) did not alter the apoptosis-inhibitory effect on ced-1(e1735) animals (Methods). Germ cell corpses were scored after 48-hour treatment of ced-1(e1735) L4 larvae. c, e, f, g, ced-1(e1735) L4 larvae were treated with UV-CM and UV-control (0.1 μg/μL) prepared from ced-3(n2433) animals ( c ), glp-1(e2141ts) animals grown at 25°C ( e ), N2 animals fed with formaldehyde-treated HB101 bacteria ( f ), and P cpr-4::cpr-4::flag; cpr-4(tm3718) animals with or without anti-Flag depletion ( g ), respectively. Data are mean ± s.e.m. The numbers of gonad arms scored are indicated inside the bars ( a–c, e–g ). ** P

    Article Snippet: 1 μL DNase (1 Unit/μL, QIAGEN), 1 μL RNase (100 μg/μL, QIAGEN) or 1 μL Trypsin (5μg/μL, Sigma) was mixed with 100 μL conditioned medium for 1 hour at 30°C.

    Techniques: Irradiation

    m 6 A alters RNA structure to recruit HNRNPG. ( A ) Sequence logo of the most enriched motif within HNRNPG PAR-CLIP peaks. ( B ) Left: secondary structure of the MALAT1 hairpin, showing the A-2,515-to-G/C/U mutations that were introduced at the m 6 A site. Right: quantification of relative HNRNPG pull-down with the original (2,515-A) and mutated (2,515-G/C/U) MALAT1 hairpins, normalized to pulled-down HIST1H1C. Data shown as mean; error bar = standard deviation; n = 3 biological replicates. ( C ) Left: structural probing of the unmethylated and methylated MALAT1 hairpins. The orange lines indicate regions with marked differences between the unmethylated and methylated hairpins. The location of the m 6 A residue is indicated by a red dot. Ctrl, no nuclease added; V1; RNase V1 digestion; S1, S1 nuclease digestion; T1, RNase T1 digestion; G-L, G-ladder; AH, alkaline hydrolysis. Right: secondary structure of the unmethylated and methylated MALAT1 hairpins, marked at their S1 nuclease (red lines) and V1 nuclease (green lines) cleavage sites. ( D ) Model showing that m 6 A disrupts RNA structure, exposes a motif that includes the m 6 A site, and recruits an RNA binding protein.

    Journal: Nucleic Acids Research

    Article Title: N6-methyladenosine alters RNA structure to regulate binding of a low-complexity protein

    doi: 10.1093/nar/gkx141

    Figure Lengend Snippet: m 6 A alters RNA structure to recruit HNRNPG. ( A ) Sequence logo of the most enriched motif within HNRNPG PAR-CLIP peaks. ( B ) Left: secondary structure of the MALAT1 hairpin, showing the A-2,515-to-G/C/U mutations that were introduced at the m 6 A site. Right: quantification of relative HNRNPG pull-down with the original (2,515-A) and mutated (2,515-G/C/U) MALAT1 hairpins, normalized to pulled-down HIST1H1C. Data shown as mean; error bar = standard deviation; n = 3 biological replicates. ( C ) Left: structural probing of the unmethylated and methylated MALAT1 hairpins. The orange lines indicate regions with marked differences between the unmethylated and methylated hairpins. The location of the m 6 A residue is indicated by a red dot. Ctrl, no nuclease added; V1; RNase V1 digestion; S1, S1 nuclease digestion; T1, RNase T1 digestion; G-L, G-ladder; AH, alkaline hydrolysis. Right: secondary structure of the unmethylated and methylated MALAT1 hairpins, marked at their S1 nuclease (red lines) and V1 nuclease (green lines) cleavage sites. ( D ) Model showing that m 6 A disrupts RNA structure, exposes a motif that includes the m 6 A site, and recruits an RNA binding protein.

    Article Snippet: The HNRNPG PAR-CLIP RNA sample was incubated with m6 A-specific antibody (202003, SYSY), RNase inhibitor (80 units, Sigma-Aldrich), human placental RNase inhibitor (NEB) in 200 μl 1× IP buffer (50 mM Tris-Cl (pH 7.4), 750 mM NaCl and 0.5% (vol/vol) Igepal CA-630) at 4°C for 2 h under gentle shaking conditions.

    Techniques: Sequencing, Cross-linking Immunoprecipitation, Standard Deviation, Methylation, RNA Binding Assay

    In vitro cell proliferation and anti-apoptotic effects of MPs. (A) Proliferation of HUVEC treated with vehicle control, with different doses of MPs or MP-treated with RNase (MP-RNase). (B) Apoptosis assay of HUVEC treated with vehicle control or with different doses of MPs. (C) Apoptosis assay of HMVEC treated with vehicle control or with different doses of MPs. Results are expressed as % compared to vehicle control and mean ± SE of three different experiments. Kruskal-Wallis test was performed; * P

    Journal: PLoS ONE

    Article Title: Microparticles from Kidney-Derived Mesenchymal Stem Cells Act as Carriers of Proangiogenic Signals and Contribute to Recovery from Acute Kidney Injury

    doi: 10.1371/journal.pone.0087853

    Figure Lengend Snippet: In vitro cell proliferation and anti-apoptotic effects of MPs. (A) Proliferation of HUVEC treated with vehicle control, with different doses of MPs or MP-treated with RNase (MP-RNase). (B) Apoptosis assay of HUVEC treated with vehicle control or with different doses of MPs. (C) Apoptosis assay of HMVEC treated with vehicle control or with different doses of MPs. Results are expressed as % compared to vehicle control and mean ± SE of three different experiments. Kruskal-Wallis test was performed; * P

    Article Snippet: To abolish the mRNA-dependent effects, MPs were pre-incubated with 1 U/ml RNase (Ambion Inc., Austin, TX, USA) for 1 h at 37°C, as previously described (RNase-treated MPs)., The reaction was stopped by adding 10 U/ml RNase inhibitor (Ambion Inc.).

    Techniques: In Vitro, Apoptosis Assay

    In vitro angiogenic effects of MPs. (A) Representative images of angiogenic effects of HUVEC pretreated with vehicle control, with different doses of MPs, or MPs preincubated with RNase (MP-RNase). (B) Quantitative analysis of tube formation of HUVEC was performed. Results are expressed as % versus vehicle alone and mean ± SEM of six different experiments. Results are expressed as mean ± SE of three different experiments. Kruskal-Wallis test was performed; * P

    Journal: PLoS ONE

    Article Title: Microparticles from Kidney-Derived Mesenchymal Stem Cells Act as Carriers of Proangiogenic Signals and Contribute to Recovery from Acute Kidney Injury

    doi: 10.1371/journal.pone.0087853

    Figure Lengend Snippet: In vitro angiogenic effects of MPs. (A) Representative images of angiogenic effects of HUVEC pretreated with vehicle control, with different doses of MPs, or MPs preincubated with RNase (MP-RNase). (B) Quantitative analysis of tube formation of HUVEC was performed. Results are expressed as % versus vehicle alone and mean ± SEM of six different experiments. Results are expressed as mean ± SE of three different experiments. Kruskal-Wallis test was performed; * P

    Article Snippet: To abolish the mRNA-dependent effects, MPs were pre-incubated with 1 U/ml RNase (Ambion Inc., Austin, TX, USA) for 1 h at 37°C, as previously described (RNase-treated MPs)., The reaction was stopped by adding 10 U/ml RNase inhibitor (Ambion Inc.).

    Techniques: In Vitro

    Anti-apoptotic effects of MPs on tubular epithelial and peritubular capillary endothelial cell in I/R injury kidney. (A) Representative confocal microscopic images of TUNEL (green) and TUNEL/CD 31 (red) double staining and (B, C) quantitative analysis of TUNEL-positive cells in the kidney 3 days after I/R injury. The total number of TUNEL-positive nuclei (B) and both TUNEL and CD 31-positive peritubular capillary nuclei (C) significantly decreased in kidneys injected with KMSCs and KMSC-derived MPs compared to those injected with vehicle control or MP-treated with RNase (MP-RNase) 3 days after IRI. Results are expressed as mean ± SE of six different experiments. Kruskal-Wallis test was performed; * P

    Journal: PLoS ONE

    Article Title: Microparticles from Kidney-Derived Mesenchymal Stem Cells Act as Carriers of Proangiogenic Signals and Contribute to Recovery from Acute Kidney Injury

    doi: 10.1371/journal.pone.0087853

    Figure Lengend Snippet: Anti-apoptotic effects of MPs on tubular epithelial and peritubular capillary endothelial cell in I/R injury kidney. (A) Representative confocal microscopic images of TUNEL (green) and TUNEL/CD 31 (red) double staining and (B, C) quantitative analysis of TUNEL-positive cells in the kidney 3 days after I/R injury. The total number of TUNEL-positive nuclei (B) and both TUNEL and CD 31-positive peritubular capillary nuclei (C) significantly decreased in kidneys injected with KMSCs and KMSC-derived MPs compared to those injected with vehicle control or MP-treated with RNase (MP-RNase) 3 days after IRI. Results are expressed as mean ± SE of six different experiments. Kruskal-Wallis test was performed; * P

    Article Snippet: To abolish the mRNA-dependent effects, MPs were pre-incubated with 1 U/ml RNase (Ambion Inc., Austin, TX, USA) for 1 h at 37°C, as previously described (RNase-treated MPs)., The reaction was stopped by adding 10 U/ml RNase inhibitor (Ambion Inc.).

    Techniques: TUNEL Assay, Double Staining, Injection, Derivative Assay

    Effects of MPs on tubular epithelial and peritubular capillary endothelial cell proliferation in I/R injury kidney. (A) Representative images of PCNA and CD 31 staining in I/R injury kidney. (B), (C) Quantitative analysis of PCNA-positive cells in kidneys 3 days after I/R injury injury. PCNA-positive counting was used to assess the proliferation of tubular epithelial cells and peritubular capillaries. There were significantly more PCNA-positive tubules (B) and both PCNA and CD 31 positive peritubular capillaries (C) in I/R injury kidneys treated with KMSC and KMSC-derived MPs compared those treated with vehicle control or MP-treated with RNase (MP-RNase) 3 days after I/R injury. Results are expressed as mean ± SE of six different experiments. Kruskal-Wallis test was performed; * P

    Journal: PLoS ONE

    Article Title: Microparticles from Kidney-Derived Mesenchymal Stem Cells Act as Carriers of Proangiogenic Signals and Contribute to Recovery from Acute Kidney Injury

    doi: 10.1371/journal.pone.0087853

    Figure Lengend Snippet: Effects of MPs on tubular epithelial and peritubular capillary endothelial cell proliferation in I/R injury kidney. (A) Representative images of PCNA and CD 31 staining in I/R injury kidney. (B), (C) Quantitative analysis of PCNA-positive cells in kidneys 3 days after I/R injury injury. PCNA-positive counting was used to assess the proliferation of tubular epithelial cells and peritubular capillaries. There were significantly more PCNA-positive tubules (B) and both PCNA and CD 31 positive peritubular capillaries (C) in I/R injury kidneys treated with KMSC and KMSC-derived MPs compared those treated with vehicle control or MP-treated with RNase (MP-RNase) 3 days after I/R injury. Results are expressed as mean ± SE of six different experiments. Kruskal-Wallis test was performed; * P

    Article Snippet: To abolish the mRNA-dependent effects, MPs were pre-incubated with 1 U/ml RNase (Ambion Inc., Austin, TX, USA) for 1 h at 37°C, as previously described (RNase-treated MPs)., The reaction was stopped by adding 10 U/ml RNase inhibitor (Ambion Inc.).

    Techniques: Staining, Derivative Assay

    Semi-quantitative injury scoring of I/R injury kidney. (A) Representative images of PAS staining in I/R injury kidney with vehicle control, KMSC, KMSC-derived MPs, and MP-treated with RNase (MP-RNase). (B) Quantitative analysis of I/R injury kidney was determined by semi-quantitative injury scoring (0–5). Injury score was significantly decreased in kidneys injected with KMSC and KMSC-derived MPs compared to those injected with vehicle control or MP-RNase 3 days after IRI. Results are expressed as % versus vehicle alone and mean ± SE of six different experiments. Kruskal-Wallis test was performed; * P

    Journal: PLoS ONE

    Article Title: Microparticles from Kidney-Derived Mesenchymal Stem Cells Act as Carriers of Proangiogenic Signals and Contribute to Recovery from Acute Kidney Injury

    doi: 10.1371/journal.pone.0087853

    Figure Lengend Snippet: Semi-quantitative injury scoring of I/R injury kidney. (A) Representative images of PAS staining in I/R injury kidney with vehicle control, KMSC, KMSC-derived MPs, and MP-treated with RNase (MP-RNase). (B) Quantitative analysis of I/R injury kidney was determined by semi-quantitative injury scoring (0–5). Injury score was significantly decreased in kidneys injected with KMSC and KMSC-derived MPs compared to those injected with vehicle control or MP-RNase 3 days after IRI. Results are expressed as % versus vehicle alone and mean ± SE of six different experiments. Kruskal-Wallis test was performed; * P

    Article Snippet: To abolish the mRNA-dependent effects, MPs were pre-incubated with 1 U/ml RNase (Ambion Inc., Austin, TX, USA) for 1 h at 37°C, as previously described (RNase-treated MPs)., The reaction was stopped by adding 10 U/ml RNase inhibitor (Ambion Inc.).

    Techniques: Staining, Derivative Assay, Injection

    Effects of MPs on microvascular rarefaction of I/R injury kidney. (A) Representative images of CD 31 stainings in I/R injury kidney with vehicle control, KMSC, KMSC-derived MPs, and MP-treated with RNase (MP-RNase). (B) Quantitative analysis of peritubular capillary rarefaction index was determined by CD31 staining. The degree of microvascular rarefaction was significantly decreased in kidneys injected with KMSC and KMSC-derived MPs compared to those injected with vehicle control or MP-RNase 3 days after I/R injury. Results are expressed as % versus vehicle alone and mean ± SE of six different experiments. Kruskal-Wallis test was performed; * P

    Journal: PLoS ONE

    Article Title: Microparticles from Kidney-Derived Mesenchymal Stem Cells Act as Carriers of Proangiogenic Signals and Contribute to Recovery from Acute Kidney Injury

    doi: 10.1371/journal.pone.0087853

    Figure Lengend Snippet: Effects of MPs on microvascular rarefaction of I/R injury kidney. (A) Representative images of CD 31 stainings in I/R injury kidney with vehicle control, KMSC, KMSC-derived MPs, and MP-treated with RNase (MP-RNase). (B) Quantitative analysis of peritubular capillary rarefaction index was determined by CD31 staining. The degree of microvascular rarefaction was significantly decreased in kidneys injected with KMSC and KMSC-derived MPs compared to those injected with vehicle control or MP-RNase 3 days after I/R injury. Results are expressed as % versus vehicle alone and mean ± SE of six different experiments. Kruskal-Wallis test was performed; * P

    Article Snippet: To abolish the mRNA-dependent effects, MPs were pre-incubated with 1 U/ml RNase (Ambion Inc., Austin, TX, USA) for 1 h at 37°C, as previously described (RNase-treated MPs)., The reaction was stopped by adding 10 U/ml RNase inhibitor (Ambion Inc.).

    Techniques: Derivative Assay, Staining, Injection