100 nt rna low molecular weight marker  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 89

    Structured Review

    Thermo Fisher 100 nt rna low molecular weight marker
    100 Nt Rna Low Molecular Weight Marker, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/100 nt rna low molecular weight marker/product/Thermo Fisher
    Average 89 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    100 nt rna low molecular weight marker - by Bioz Stars, 2020-04
    89/100 stars

    Related Products / Commonly Used Together

    rna substrates
    polyacrylamide gel
    γ32 p atp-labeled 17 -- 25

    Images

    Related Articles

    In Vitro:

    Article Title: Nucleotide sequence of miRNA precursor contributes to cleavage site selection by Dicer
    Article Snippet: Paragraph title: Dicer in vitro cleavage products visualized by northern blotting ... Briefly, the RNA substrates and products (25 ng) were separated on a 12% denaturing polyacrylamide gel along with a [γ32 P]ATP-labeled 17–25 nt RNA marker, a labeled 10- to 100-nt RNA Low Molecular Weight Marker (USB Corp.) and appropriate phosphorylated M5 and M3 markers (indicated in the figure legends) (Supplementary Table S2).

    Northern Blot:

    Article Title: Nucleotide sequence of miRNA precursor contributes to cleavage site selection by Dicer
    Article Snippet: Paragraph title: Dicer in vitro cleavage products visualized by northern blotting ... Briefly, the RNA substrates and products (25 ng) were separated on a 12% denaturing polyacrylamide gel along with a [γ32 P]ATP-labeled 17–25 nt RNA marker, a labeled 10- to 100-nt RNA Low Molecular Weight Marker (USB Corp.) and appropriate phosphorylated M5 and M3 markers (indicated in the figure legends) (Supplementary Table S2).

    Labeling:

    Article Title: Nucleotide sequence of miRNA precursor contributes to cleavage site selection by Dicer
    Article Snippet: .. Briefly, the RNA substrates and products (25 ng) were separated on a 12% denaturing polyacrylamide gel along with a [γ32 P]ATP-labeled 17–25 nt RNA marker, a labeled 10- to 100-nt RNA Low Molecular Weight Marker (USB Corp.) and appropriate phosphorylated M5 and M3 markers (indicated in the figure legends) (Supplementary Table S2). .. After electrophoresis, the RNAs were transferred to a GeneScreen Plus hybridization membrane (Perkin Elmer).

    Electrophoresis:

    Article Title: Nucleotide sequence of miRNA precursor contributes to cleavage site selection by Dicer
    Article Snippet: Briefly, the RNA substrates and products (25 ng) were separated on a 12% denaturing polyacrylamide gel along with a [γ32 P]ATP-labeled 17–25 nt RNA marker, a labeled 10- to 100-nt RNA Low Molecular Weight Marker (USB Corp.) and appropriate phosphorylated M5 and M3 markers (indicated in the figure legends) (Supplementary Table S2). .. After electrophoresis, the RNAs were transferred to a GeneScreen Plus hybridization membrane (Perkin Elmer).

    Immunoprecipitation:

    Article Title: Nucleotide sequence of miRNA precursor contributes to cleavage site selection by Dicer
    Article Snippet: Dicer in vitro cleavage products visualized by northern blotting The phosphorylated RNAs (50 ng, (∼2.5 pmole)) were cleaved with 0.75 U (0.187 pmole) of Dicer (Genlantis) for 12 min at 37°C, as previously described ( , ) or with 2 μl of immunoprecipitated (10% of total pull-down eluate volume) by the pull-down Dicer–TRBP complex for 15 min at 37°C. .. Briefly, the RNA substrates and products (25 ng) were separated on a 12% denaturing polyacrylamide gel along with a [γ32 P]ATP-labeled 17–25 nt RNA marker, a labeled 10- to 100-nt RNA Low Molecular Weight Marker (USB Corp.) and appropriate phosphorylated M5 and M3 markers (indicated in the figure legends) (Supplementary Table S2).

    Marker:

    Article Title: Nucleotide sequence of miRNA precursor contributes to cleavage site selection by Dicer
    Article Snippet: .. Briefly, the RNA substrates and products (25 ng) were separated on a 12% denaturing polyacrylamide gel along with a [γ32 P]ATP-labeled 17–25 nt RNA marker, a labeled 10- to 100-nt RNA Low Molecular Weight Marker (USB Corp.) and appropriate phosphorylated M5 and M3 markers (indicated in the figure legends) (Supplementary Table S2). .. After electrophoresis, the RNAs were transferred to a GeneScreen Plus hybridization membrane (Perkin Elmer).

    Molecular Weight:

    Article Title: Nucleotide sequence of miRNA precursor contributes to cleavage site selection by Dicer
    Article Snippet: .. Briefly, the RNA substrates and products (25 ng) were separated on a 12% denaturing polyacrylamide gel along with a [γ32 P]ATP-labeled 17–25 nt RNA marker, a labeled 10- to 100-nt RNA Low Molecular Weight Marker (USB Corp.) and appropriate phosphorylated M5 and M3 markers (indicated in the figure legends) (Supplementary Table S2). .. After electrophoresis, the RNAs were transferred to a GeneScreen Plus hybridization membrane (Perkin Elmer).

    Hybridization:

    Article Title: Nucleotide sequence of miRNA precursor contributes to cleavage site selection by Dicer
    Article Snippet: Briefly, the RNA substrates and products (25 ng) were separated on a 12% denaturing polyacrylamide gel along with a [γ32 P]ATP-labeled 17–25 nt RNA marker, a labeled 10- to 100-nt RNA Low Molecular Weight Marker (USB Corp.) and appropriate phosphorylated M5 and M3 markers (indicated in the figure legends) (Supplementary Table S2). .. After electrophoresis, the RNAs were transferred to a GeneScreen Plus hybridization membrane (Perkin Elmer).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher sirna
    Suppression of Akt and PKC signaling contributes to <t>INPP4B</t> regulation of AR transcriptional activity. (A) VCaP cells were transfected with control, INPP4B, or PTEN specific siRNAs. Expression levels of INPP4B, PTEN, pPKCζ, pPKCβII and β-tubulin were measured by Western blotting. (B) LNCaP cells were transfected with control or INPP4B siRNAs for 48 hours in medium supplemented with 10% FBS. Cellular lysates were assayed for INPP4B, pAkt, pPKCζ, pPKCβII and β-tubulin by Western blotting. (C) C4–2 cells were transfected and assayed in parallel with B. (D) LNCaP cells were plated in complete medium and treated with indicated inhibitors for 8 hours. Protein extracts were assayed for AR, INPP4B, pS6, and tubulin levels by Western blotting. (E-J) LNCaP cells were plated in medium supplemented with 10% CSS with either vehicle or 1 nM R1881. Twenty four hours later cells were treated with vehicle (DMSO), 5μM AZD5363, or 2 μM BIM-I for an additional 8 hours. In parallel, LNCaP cells were transfected with control or INPP4B <t>siRNA</t> for 48 hours in 10% CSS medium supplemented with 1 nM R1881. RNA was purified, reverse transcribed, and expression levels of AR regulated genes PLA2G2A (E), KIAA1324 (F), TARP (G), NNMT (H), TMPRSS2 (I) and SYTL2 (J) compared by RT-qPCR. ** represents p
    Sirna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2186 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirna/product/Thermo Fisher
    Average 99 stars, based on 2186 article reviews
    Price from $9.99 to $1999.99
    sirna - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    98
    Thermo Fisher rna
    T- or A-tracts, adjacent to −35 elements, regulate gene expression by a rheostat-like mechanism. Schematic overview of the T-tract rheostat using the sabA promoter as a model. The predicted interaction of the <t>RNA</t> polymerase with sabA promoter, harboring different T-tract lengths and thereby different local <t>DNA</t> structure, is depicted in the model. The illustration shows the high-expressing T 13 -variant and the low-expressing T 18 -variant. The region containing the A-boxes, i.e. the proximal UP-like element, is marked in purple (−90 to −50), T-tract in blue, and the core promoter (−35 to +20) in yellow. Bent arrows indicate the change in local DNA structure that occurs in two orientations as the T-tract length is altered. This is a variable process as the T-tract length can both be lengthened and shortened, as a result of slipped strand mispairing during replication.
    Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 3930 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rna/product/Thermo Fisher
    Average 98 stars, based on 3930 article reviews
    Price from $9.99 to $1999.99
    rna - by Bioz Stars, 2020-04
    98/100 stars
      Buy from Supplier

    Image Search Results


    Suppression of Akt and PKC signaling contributes to INPP4B regulation of AR transcriptional activity. (A) VCaP cells were transfected with control, INPP4B, or PTEN specific siRNAs. Expression levels of INPP4B, PTEN, pPKCζ, pPKCβII and β-tubulin were measured by Western blotting. (B) LNCaP cells were transfected with control or INPP4B siRNAs for 48 hours in medium supplemented with 10% FBS. Cellular lysates were assayed for INPP4B, pAkt, pPKCζ, pPKCβII and β-tubulin by Western blotting. (C) C4–2 cells were transfected and assayed in parallel with B. (D) LNCaP cells were plated in complete medium and treated with indicated inhibitors for 8 hours. Protein extracts were assayed for AR, INPP4B, pS6, and tubulin levels by Western blotting. (E-J) LNCaP cells were plated in medium supplemented with 10% CSS with either vehicle or 1 nM R1881. Twenty four hours later cells were treated with vehicle (DMSO), 5μM AZD5363, or 2 μM BIM-I for an additional 8 hours. In parallel, LNCaP cells were transfected with control or INPP4B siRNA for 48 hours in 10% CSS medium supplemented with 1 nM R1881. RNA was purified, reverse transcribed, and expression levels of AR regulated genes PLA2G2A (E), KIAA1324 (F), TARP (G), NNMT (H), TMPRSS2 (I) and SYTL2 (J) compared by RT-qPCR. ** represents p

    Journal: Oncogene

    Article Title: Inositol Polyphosphate 4-phosphatase Type II Regulation of Androgen Receptor Activity

    doi: 10.1038/s41388-018-0498-3

    Figure Lengend Snippet: Suppression of Akt and PKC signaling contributes to INPP4B regulation of AR transcriptional activity. (A) VCaP cells were transfected with control, INPP4B, or PTEN specific siRNAs. Expression levels of INPP4B, PTEN, pPKCζ, pPKCβII and β-tubulin were measured by Western blotting. (B) LNCaP cells were transfected with control or INPP4B siRNAs for 48 hours in medium supplemented with 10% FBS. Cellular lysates were assayed for INPP4B, pAkt, pPKCζ, pPKCβII and β-tubulin by Western blotting. (C) C4–2 cells were transfected and assayed in parallel with B. (D) LNCaP cells were plated in complete medium and treated with indicated inhibitors for 8 hours. Protein extracts were assayed for AR, INPP4B, pS6, and tubulin levels by Western blotting. (E-J) LNCaP cells were plated in medium supplemented with 10% CSS with either vehicle or 1 nM R1881. Twenty four hours later cells were treated with vehicle (DMSO), 5μM AZD5363, or 2 μM BIM-I for an additional 8 hours. In parallel, LNCaP cells were transfected with control or INPP4B siRNA for 48 hours in 10% CSS medium supplemented with 1 nM R1881. RNA was purified, reverse transcribed, and expression levels of AR regulated genes PLA2G2A (E), KIAA1324 (F), TARP (G), NNMT (H), TMPRSS2 (I) and SYTL2 (J) compared by RT-qPCR. ** represents p

    Article Snippet: siRNA transfections Noncoding control, INPP4B, PTEN, and FOXA1 siRNAs were transfected at 50–100 pmol siRNA per well in 6 well cell culture plate using Lipofectamine RNAiMAX Reagent (Thermo Fisher Scientific) as recommended by the manufacturer.

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

    INPP4B transcription is regulated by full length AR and FOXA1. LNCaP cells in 10% CSS were transfected with noncoding control or FOXA1 specific siRNA; cells were treated with R1881 after 24 hours. Cells were harvested after 48 hours and assayed for FOXA1 expression by western blotting (A), RNA isolated and assayed for FKBP5 (B), RASSF3 (C), and INPP4B (D). ( E) A diagram of AR, FOXA1, and Pol2 recruitment and RNA-seq in LNCaP cells. (F-J) LNCaP AR-V7/pHage cells were placed in medium with 10% CSS for 24 hours and treated with 50 ng/ml Dox or 10 nM R1881 overnight as indicated. Recruitment of AR-FL and AR-V7 to PSA enhancer (F), INPP4B intron 2 (G), INPP4B enhancer region (H), INPP4B transcription start site chr4:143522349 (I), and chr4:143514774 (J) were measured by ChIP-qPCR using Igg as control. * p

    Journal: Oncogene

    Article Title: Inositol Polyphosphate 4-phosphatase Type II Regulation of Androgen Receptor Activity

    doi: 10.1038/s41388-018-0498-3

    Figure Lengend Snippet: INPP4B transcription is regulated by full length AR and FOXA1. LNCaP cells in 10% CSS were transfected with noncoding control or FOXA1 specific siRNA; cells were treated with R1881 after 24 hours. Cells were harvested after 48 hours and assayed for FOXA1 expression by western blotting (A), RNA isolated and assayed for FKBP5 (B), RASSF3 (C), and INPP4B (D). ( E) A diagram of AR, FOXA1, and Pol2 recruitment and RNA-seq in LNCaP cells. (F-J) LNCaP AR-V7/pHage cells were placed in medium with 10% CSS for 24 hours and treated with 50 ng/ml Dox or 10 nM R1881 overnight as indicated. Recruitment of AR-FL and AR-V7 to PSA enhancer (F), INPP4B intron 2 (G), INPP4B enhancer region (H), INPP4B transcription start site chr4:143522349 (I), and chr4:143514774 (J) were measured by ChIP-qPCR using Igg as control. * p

    Article Snippet: siRNA transfections Noncoding control, INPP4B, PTEN, and FOXA1 siRNAs were transfected at 50–100 pmol siRNA per well in 6 well cell culture plate using Lipofectamine RNAiMAX Reagent (Thermo Fisher Scientific) as recommended by the manufacturer.

    Techniques: Transfection, Expressing, Western Blot, Isolation, RNA Sequencing Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction

    Reciprocal regulation of AR-V7 and INPP4B. (A) LNCaP AR-V7/pHage cells were placed in medium with 10% CSS or 10% TET FBS serum for 24 hours and treated with 50 ng/ml Dox or 1 nM R1881 as indicated for an additional 36 hours. Protein level of INPP4B, AR-FL, AR-V7, pAkt, pPKCβII, and tubulin were assessed by Western blotting. (B, C) LNCaP AR-V7/pHage cells were transfected with control or INPP4B siRNA and treated with either vehicle or 25 ng/ml Dox for additional 24 hours. Gene expression of INPP4B (B) and EDN2 (C) were analyzed by RT-qPCR using 18S as a control. (D-I) LNCaP AR-V7/pHage cells were transfected and treated as in B. RNA was analyzed for expression of NNMT (D), PSA (E), TARP (F), TMPRSS2 (G), KIAA1324 (H), and SYTL2 (I).*** p

    Journal: Oncogene

    Article Title: Inositol Polyphosphate 4-phosphatase Type II Regulation of Androgen Receptor Activity

    doi: 10.1038/s41388-018-0498-3

    Figure Lengend Snippet: Reciprocal regulation of AR-V7 and INPP4B. (A) LNCaP AR-V7/pHage cells were placed in medium with 10% CSS or 10% TET FBS serum for 24 hours and treated with 50 ng/ml Dox or 1 nM R1881 as indicated for an additional 36 hours. Protein level of INPP4B, AR-FL, AR-V7, pAkt, pPKCβII, and tubulin were assessed by Western blotting. (B, C) LNCaP AR-V7/pHage cells were transfected with control or INPP4B siRNA and treated with either vehicle or 25 ng/ml Dox for additional 24 hours. Gene expression of INPP4B (B) and EDN2 (C) were analyzed by RT-qPCR using 18S as a control. (D-I) LNCaP AR-V7/pHage cells were transfected and treated as in B. RNA was analyzed for expression of NNMT (D), PSA (E), TARP (F), TMPRSS2 (G), KIAA1324 (H), and SYTL2 (I).*** p

    Article Snippet: siRNA transfections Noncoding control, INPP4B, PTEN, and FOXA1 siRNAs were transfected at 50–100 pmol siRNA per well in 6 well cell culture plate using Lipofectamine RNAiMAX Reagent (Thermo Fisher Scientific) as recommended by the manufacturer.

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

    siRNAs targeted to mouse IGF-IR in breast cancer cells. ( A ) The effect of siRNAs (10 nM) designed against IGF-IR coding regions was measured by qRT-PCR 24 h post-transfection in EMT6 cells. ( B ) Dose-dependent inhibition of IGF-IR mRNA levels by ADT siRNA into EMT6. ( C ) Kinetics of silencing by ADT siRNA in EMT6 cells. Black bars represent mock transfected cells (NT). Means ± SEM of two independent experiments with independent transfected quadruplicates were shown; * P

    Journal: PLoS ONE

    Article Title: Small Interfering RNA Targeted to IGF-IR Delays Tumor Growth and Induces Proinflammatory Cytokines in a Mouse Breast Cancer Model

    doi: 10.1371/journal.pone.0029213

    Figure Lengend Snippet: siRNAs targeted to mouse IGF-IR in breast cancer cells. ( A ) The effect of siRNAs (10 nM) designed against IGF-IR coding regions was measured by qRT-PCR 24 h post-transfection in EMT6 cells. ( B ) Dose-dependent inhibition of IGF-IR mRNA levels by ADT siRNA into EMT6. ( C ) Kinetics of silencing by ADT siRNA in EMT6 cells. Black bars represent mock transfected cells (NT). Means ± SEM of two independent experiments with independent transfected quadruplicates were shown; * P

    Article Snippet: In vivo tumor growth experiments C4HD cells growing in MPA (10 nM) were transiently transfected with 2′-O-methyl- modified siRNA (100 nM) using Dharmafect-I (Thermo Fisher Scientific, USA).

    Techniques: Quantitative RT-PCR, Transfection, Inhibition

    Immunization with C4HD cells treated with IGF-IR siRNAs. ( A ) Delayed-type hypersensitivity (DTH) response. BALB/c mice (n = 10) were immunized with three injections of irradiated C4HD cells (black bar) or irradiated 2′-O-methyl ADT siRNA (white bar) or 2′-O-methyl CONT2 siRNA (grey bar) transfected C4HD. NT, non-transfected cells. Data are presented as mean ± SEM; *** P

    Journal: PLoS ONE

    Article Title: Small Interfering RNA Targeted to IGF-IR Delays Tumor Growth and Induces Proinflammatory Cytokines in a Mouse Breast Cancer Model

    doi: 10.1371/journal.pone.0029213

    Figure Lengend Snippet: Immunization with C4HD cells treated with IGF-IR siRNAs. ( A ) Delayed-type hypersensitivity (DTH) response. BALB/c mice (n = 10) were immunized with three injections of irradiated C4HD cells (black bar) or irradiated 2′-O-methyl ADT siRNA (white bar) or 2′-O-methyl CONT2 siRNA (grey bar) transfected C4HD. NT, non-transfected cells. Data are presented as mean ± SEM; *** P

    Article Snippet: In vivo tumor growth experiments C4HD cells growing in MPA (10 nM) were transiently transfected with 2′-O-methyl- modified siRNA (100 nM) using Dharmafect-I (Thermo Fisher Scientific, USA).

    Techniques: Mouse Assay, Irradiation, Transfection

    IGF-IR gene silencing inhibits signaling, cell cycle progression and proliferation. EMT6 cells were transfected with 25 to 100 nM 2′-O-methyl ADT siRNA and harvested 48 h later after standard growth conditions (10% FCS) for analysis by immunoblotting with: ( A ) antibodies against phospho-S473 AKT and total AKT, and ( B ) antibodies against phospho-T202/T204 ERKs and total ERK. Black bars represent mock transfected cells (NT). Representative experiments are presented and quantitative analysis of three independent experiments are shown on the right of each gel with means ± SEM. * P

    Journal: PLoS ONE

    Article Title: Small Interfering RNA Targeted to IGF-IR Delays Tumor Growth and Induces Proinflammatory Cytokines in a Mouse Breast Cancer Model

    doi: 10.1371/journal.pone.0029213

    Figure Lengend Snippet: IGF-IR gene silencing inhibits signaling, cell cycle progression and proliferation. EMT6 cells were transfected with 25 to 100 nM 2′-O-methyl ADT siRNA and harvested 48 h later after standard growth conditions (10% FCS) for analysis by immunoblotting with: ( A ) antibodies against phospho-S473 AKT and total AKT, and ( B ) antibodies against phospho-T202/T204 ERKs and total ERK. Black bars represent mock transfected cells (NT). Representative experiments are presented and quantitative analysis of three independent experiments are shown on the right of each gel with means ± SEM. * P

    Article Snippet: In vivo tumor growth experiments C4HD cells growing in MPA (10 nM) were transiently transfected with 2′-O-methyl- modified siRNA (100 nM) using Dharmafect-I (Thermo Fisher Scientific, USA).

    Techniques: Transfection

    T- or A-tracts, adjacent to −35 elements, regulate gene expression by a rheostat-like mechanism. Schematic overview of the T-tract rheostat using the sabA promoter as a model. The predicted interaction of the RNA polymerase with sabA promoter, harboring different T-tract lengths and thereby different local DNA structure, is depicted in the model. The illustration shows the high-expressing T 13 -variant and the low-expressing T 18 -variant. The region containing the A-boxes, i.e. the proximal UP-like element, is marked in purple (−90 to −50), T-tract in blue, and the core promoter (−35 to +20) in yellow. Bent arrows indicate the change in local DNA structure that occurs in two orientations as the T-tract length is altered. This is a variable process as the T-tract length can both be lengthened and shortened, as a result of slipped strand mispairing during replication.

    Journal: PLoS Pathogens

    Article Title: A Repetitive DNA Element Regulates Expression of the Helicobacter pylori Sialic Acid Binding Adhesin by a Rheostat-like Mechanism

    doi: 10.1371/journal.ppat.1004234

    Figure Lengend Snippet: T- or A-tracts, adjacent to −35 elements, regulate gene expression by a rheostat-like mechanism. Schematic overview of the T-tract rheostat using the sabA promoter as a model. The predicted interaction of the RNA polymerase with sabA promoter, harboring different T-tract lengths and thereby different local DNA structure, is depicted in the model. The illustration shows the high-expressing T 13 -variant and the low-expressing T 18 -variant. The region containing the A-boxes, i.e. the proximal UP-like element, is marked in purple (−90 to −50), T-tract in blue, and the core promoter (−35 to +20) in yellow. Bent arrows indicate the change in local DNA structure that occurs in two orientations as the T-tract length is altered. This is a variable process as the T-tract length can both be lengthened and shortened, as a result of slipped strand mispairing during replication.

    Article Snippet: 20 µl reactions, using cDNA from 100 ng of RNA as template, Phusion Hot Start DNA polymerase (Thermo Scientific) and gene specific primers (5 µM each), were run on MJ PTC-200 thermal cycler (MJ Research).

    Techniques: Expressing, Variant Assay

    Effect of curcumin and curcumin A on HIV-1 messenger (m)RNA expression, HIV-1 transcription and HIV-1 reverse transcription. Notes: ( A ) Effect of curcumin A on HIV-1 mRNA expr ession. CEM-T cells infected with HIV-1 Luc were treated with DMSO, 1 μM curcumin or 1 μM curcumin A for 48 hours as indicated. RNA was extracted, reverse transcribed, and analyzed with primers for HIV-1 gag and env genes by real-time PCR on Roche 4800 (Hoffman-La Roche Ltd., Basel, Switzerland) using 18S RNA as a reference. ( B , C ). Effect of curcumin and curcumin A on Tat-induced and basal HIV-1 transcription. In panel ( B ) 293T cells were transiently transfected with a vector contacting HIV LTR followed by the luciferase reporter (HIV LTR) and Tat expression vector or pNL4-3.Luc. In panel ( C ) 293T cells were transfected with HIV-1 LTR expression vector or with HIV LTR with the inactivated SP1 sites (NF-κB). For normalization, the cells were also co-transfected with GFP expressing vector. At 24 hours post-transfection the cells were treated with 1 μM curcumin or curcumin A for 24 hours. Then the cells were lyzed and luciferase activity was measured. GFP fluorescence was measured in parallel and used for normalization. ( D ) Effect of curcumin and curcumin A on HIV-1 reverse transcription. CEM-T cells were infected with HIV-1 Luc and then treated with DMSO, AZT, 1 μM curcumin or curcumin A as indicated for 6 hours. DNA was extracted and analyzed by real-time PCR on Roche 4800 using primers for early and late LTR and β-globin gene as a reference. Abbreviations: DMSO, dimethyl sulfoxide; mRNA, messenger RNA; PCR, polymerase chain reaction; AZT, azidothymidine.

    Journal: Drug Design, Development and Therapy

    Article Title: Inhibition of HIV-1 by curcumin A, a novel curcumin analog

    doi: 10.2147/DDDT.S86558

    Figure Lengend Snippet: Effect of curcumin and curcumin A on HIV-1 messenger (m)RNA expression, HIV-1 transcription and HIV-1 reverse transcription. Notes: ( A ) Effect of curcumin A on HIV-1 mRNA expr ession. CEM-T cells infected with HIV-1 Luc were treated with DMSO, 1 μM curcumin or 1 μM curcumin A for 48 hours as indicated. RNA was extracted, reverse transcribed, and analyzed with primers for HIV-1 gag and env genes by real-time PCR on Roche 4800 (Hoffman-La Roche Ltd., Basel, Switzerland) using 18S RNA as a reference. ( B , C ). Effect of curcumin and curcumin A on Tat-induced and basal HIV-1 transcription. In panel ( B ) 293T cells were transiently transfected with a vector contacting HIV LTR followed by the luciferase reporter (HIV LTR) and Tat expression vector or pNL4-3.Luc. In panel ( C ) 293T cells were transfected with HIV-1 LTR expression vector or with HIV LTR with the inactivated SP1 sites (NF-κB). For normalization, the cells were also co-transfected with GFP expressing vector. At 24 hours post-transfection the cells were treated with 1 μM curcumin or curcumin A for 24 hours. Then the cells were lyzed and luciferase activity was measured. GFP fluorescence was measured in parallel and used for normalization. ( D ) Effect of curcumin and curcumin A on HIV-1 reverse transcription. CEM-T cells were infected with HIV-1 Luc and then treated with DMSO, AZT, 1 μM curcumin or curcumin A as indicated for 6 hours. DNA was extracted and analyzed by real-time PCR on Roche 4800 using primers for early and late LTR and β-globin gene as a reference. Abbreviations: DMSO, dimethyl sulfoxide; mRNA, messenger RNA; PCR, polymerase chain reaction; AZT, azidothymidine.

    Article Snippet: Total RNA (100 ng) was reverse-transcribed to complementary (c)DNA using Superscript™ RT-PCR (reverse transcription polymerase chain reaction) kit (Thermo Fisher Scientific); hexamers and oligo-dT were used as primers.

    Techniques: Expressing, Infection, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Fluorescence, Polymerase Chain Reaction