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Promega oligo dt primers
MiR‐24 facilitates non‐small cell lung cancer (NSCLC) cell growth in a tumor xenograft model. ( a ) An equal number of NCI‐H358 cells was injected into two groups of nude mice treated with miR‐24 inhibitor or scrambled <t>oligo</t> through local injection of the xenograft tumor every five days, seven days after the injection. The image shows the tumor xenograft. ( b ) Tumor size was measured every three days, after seven days of injections. The tumor volume was calculated as follows: length × width 2 × 1/2. ( ) Ctrl and ( ) miR‐24 inhibitor. ( c ) miR‐24 and ( d ) <t>WWOX</t> expression in the xenograft tumor. ( e ) c‐Kit expression in the xenograft tumor by immunohistochemistry. Error bars indicate the mean ± standard deviation of three independent experiments, * P
Oligo Dt Primers, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 241 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Inhibition of miR‐24 suppresses malignancy of human non‐small cell lung cancer cells by targeting WWOX in vitro and in vivo"

Article Title: Inhibition of miR‐24 suppresses malignancy of human non‐small cell lung cancer cells by targeting WWOX in vitro and in vivo

Journal: Thoracic Cancer

doi: 10.1111/1759-7714.12824

MiR‐24 facilitates non‐small cell lung cancer (NSCLC) cell growth in a tumor xenograft model. ( a ) An equal number of NCI‐H358 cells was injected into two groups of nude mice treated with miR‐24 inhibitor or scrambled oligo through local injection of the xenograft tumor every five days, seven days after the injection. The image shows the tumor xenograft. ( b ) Tumor size was measured every three days, after seven days of injections. The tumor volume was calculated as follows: length × width 2 × 1/2. ( ) Ctrl and ( ) miR‐24 inhibitor. ( c ) miR‐24 and ( d ) WWOX expression in the xenograft tumor. ( e ) c‐Kit expression in the xenograft tumor by immunohistochemistry. Error bars indicate the mean ± standard deviation of three independent experiments, * P
Figure Legend Snippet: MiR‐24 facilitates non‐small cell lung cancer (NSCLC) cell growth in a tumor xenograft model. ( a ) An equal number of NCI‐H358 cells was injected into two groups of nude mice treated with miR‐24 inhibitor or scrambled oligo through local injection of the xenograft tumor every five days, seven days after the injection. The image shows the tumor xenograft. ( b ) Tumor size was measured every three days, after seven days of injections. The tumor volume was calculated as follows: length × width 2 × 1/2. ( ) Ctrl and ( ) miR‐24 inhibitor. ( c ) miR‐24 and ( d ) WWOX expression in the xenograft tumor. ( e ) c‐Kit expression in the xenograft tumor by immunohistochemistry. Error bars indicate the mean ± standard deviation of three independent experiments, * P

Techniques Used: Injection, Mouse Assay, Expressing, Immunohistochemistry, Standard Deviation

2) Product Images from "Inhibition of miR‐24 suppresses malignancy of human non‐small cell lung cancer cells by targeting WWOX in vitro and in vivo"

Article Title: Inhibition of miR‐24 suppresses malignancy of human non‐small cell lung cancer cells by targeting WWOX in vitro and in vivo

Journal: Thoracic Cancer

doi: 10.1111/1759-7714.12824

MiR‐24 facilitates non‐small cell lung cancer (NSCLC) cell growth in a tumor xenograft model. ( a ) An equal number of NCI‐H358 cells was injected into two groups of nude mice treated with miR‐24 inhibitor or scrambled oligo through local injection of the xenograft tumor every five days, seven days after the injection. The image shows the tumor xenograft. ( b ) Tumor size was measured every three days, after seven days of injections. The tumor volume was calculated as follows: length × width 2 × 1/2. ( ) Ctrl and ( ) miR‐24 inhibitor. ( c ) miR‐24 and ( d ) WWOX expression in the xenograft tumor. ( e ) c‐Kit expression in the xenograft tumor by immunohistochemistry. Error bars indicate the mean ± standard deviation of three independent experiments, * P
Figure Legend Snippet: MiR‐24 facilitates non‐small cell lung cancer (NSCLC) cell growth in a tumor xenograft model. ( a ) An equal number of NCI‐H358 cells was injected into two groups of nude mice treated with miR‐24 inhibitor or scrambled oligo through local injection of the xenograft tumor every five days, seven days after the injection. The image shows the tumor xenograft. ( b ) Tumor size was measured every three days, after seven days of injections. The tumor volume was calculated as follows: length × width 2 × 1/2. ( ) Ctrl and ( ) miR‐24 inhibitor. ( c ) miR‐24 and ( d ) WWOX expression in the xenograft tumor. ( e ) c‐Kit expression in the xenograft tumor by immunohistochemistry. Error bars indicate the mean ± standard deviation of three independent experiments, * P

Techniques Used: Injection, Mouse Assay, Expressing, Immunohistochemistry, Standard Deviation

3) Product Images from "Drosophila p120catenin plays a supporting role in cell adhesion but is not an essential adherens junction component"

Article Title: Drosophila p120catenin plays a supporting role in cell adhesion but is not an essential adherens junction component

Journal: The Journal of Cell Biology

doi: 10.1083/jcb.200211083

p120 308 is a null allele. (A) p120 308 deletes the entire p120 coding region, but does not affect other genes. Schematic as in Fig. 1 A. Genomic DNA from single wild-type or homozygous mutant flies was PCR amplified using primer pairs from the indicated regions between LD05623 and CG17486 . (B) The p120 mutants are mRNA nulls. cDNA generated from oligo-dT-primed total RNA from p120 mutants and wild-type was amplified with primers spanning the p120 third intron. An unrelated gene, CG2905 , is a control. A DNA control confirmed we were examining mRNA.
Figure Legend Snippet: p120 308 is a null allele. (A) p120 308 deletes the entire p120 coding region, but does not affect other genes. Schematic as in Fig. 1 A. Genomic DNA from single wild-type or homozygous mutant flies was PCR amplified using primer pairs from the indicated regions between LD05623 and CG17486 . (B) The p120 mutants are mRNA nulls. cDNA generated from oligo-dT-primed total RNA from p120 mutants and wild-type was amplified with primers spanning the p120 third intron. An unrelated gene, CG2905 , is a control. A DNA control confirmed we were examining mRNA.

Techniques Used: Mutagenesis, Polymerase Chain Reaction, Amplification, Generated

4) Product Images from "p100 Deficiency Is Insufficient for Full Activation of the Alternative NF-?B Pathway: TNF Cooperates with p52-RelB in Target Gene Transcription"

Article Title: p100 Deficiency Is Insufficient for Full Activation of the Alternative NF-?B Pathway: TNF Cooperates with p52-RelB in Target Gene Transcription

Journal: PLoS ONE

doi: 10.1371/journal.pone.0042741

Expression profiling of wild-type and p100 −/− MEFs. (A) Western blot analysis of cytoplasmic and nuclear protein extracts (20 µg/sample) were analyzed for the presence of NF-κB family members p100/p52, RelB, and RelA in wild-type (+/+) and in p100 −/− (−/−) MEFs. As cytoplasmic loading control S6 ribosomal protein and as nuclear loading control RNA Pol II was assayed. (B) Increased κB DNA-binding activity in nuclear extracts from p100 −/− (−/−) compared to wild-type MEFs (+/+). Five µg protein extract per cell line were incubated with a radioactively labeled Igκ oligo and analyzed by EMSA. Supershift analysis was performed using pre-immune serum (p.i.), anti-RelA (α-RelA), anti-RelB (α-RelB), and anti-p52 antibodies (α-p52). Super-shifted RelB and p52 complexes are indicated by arrow and arrowhead, respectively. (C) Heatmap displaying fold change values observed in p100 −/− vs. wild-type cells. The color code indicates the fold change values between +6 fold up- (red) and −20 fold downregulation (green). Each horizontal line on the heatmap corresponds to one gene. Genes labeled with blue boxes on the left were verified by qRT-PCR. Gene symbols and abbreviations of GO terms are displayed on the right. CA, Cytokine activity; GPCRB, G-protein-coupled receptor binding; IGFB, insulin-like growth factor binding; ER, extracellular region; ESO, extracellular structure organization and biogenesis; D/M, developmental process; B/ND, forebrain development and nervous system development; CG/S, regulation of cell size; IR/RES, immune response and response to external stimulus; M/L/T, locomotory behavior. (D) Significantly regulated Gene Ontology terms with respective gene numbers.
Figure Legend Snippet: Expression profiling of wild-type and p100 −/− MEFs. (A) Western blot analysis of cytoplasmic and nuclear protein extracts (20 µg/sample) were analyzed for the presence of NF-κB family members p100/p52, RelB, and RelA in wild-type (+/+) and in p100 −/− (−/−) MEFs. As cytoplasmic loading control S6 ribosomal protein and as nuclear loading control RNA Pol II was assayed. (B) Increased κB DNA-binding activity in nuclear extracts from p100 −/− (−/−) compared to wild-type MEFs (+/+). Five µg protein extract per cell line were incubated with a radioactively labeled Igκ oligo and analyzed by EMSA. Supershift analysis was performed using pre-immune serum (p.i.), anti-RelA (α-RelA), anti-RelB (α-RelB), and anti-p52 antibodies (α-p52). Super-shifted RelB and p52 complexes are indicated by arrow and arrowhead, respectively. (C) Heatmap displaying fold change values observed in p100 −/− vs. wild-type cells. The color code indicates the fold change values between +6 fold up- (red) and −20 fold downregulation (green). Each horizontal line on the heatmap corresponds to one gene. Genes labeled with blue boxes on the left were verified by qRT-PCR. Gene symbols and abbreviations of GO terms are displayed on the right. CA, Cytokine activity; GPCRB, G-protein-coupled receptor binding; IGFB, insulin-like growth factor binding; ER, extracellular region; ESO, extracellular structure organization and biogenesis; D/M, developmental process; B/ND, forebrain development and nervous system development; CG/S, regulation of cell size; IR/RES, immune response and response to external stimulus; M/L/T, locomotory behavior. (D) Significantly regulated Gene Ontology terms with respective gene numbers.

Techniques Used: Expressing, Western Blot, Binding Assay, Activity Assay, Incubation, Labeling, Quantitative RT-PCR

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Amplification:

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Article Snippet: .. Reverse transcription-PCR (RT-PCR) and quantitative real-time RT-PCR Total RNA was reverse-transcribed and amplified using reverse transcription and PCR kits, respectively (Promega Corp., Madison, WI, U.S.). .. Real-time RT-PCR was performed by Bio-Rad IQ5 (Bio-Rad, U.S.).

Quantitative RT-PCR:

Article Title: Effects of THAP11 on Erythroid Differentiation and Megakaryocytic Differentiation of K562 Cells
Article Snippet: .. Reverse transcription-PCR (RT-PCR) and quantitative real-time RT-PCR Total RNA was reverse-transcribed and amplified using reverse transcription and PCR kits, respectively (Promega Corp., Madison, WI, U.S.). .. Real-time RT-PCR was performed by Bio-Rad IQ5 (Bio-Rad, U.S.).

Real-time Polymerase Chain Reaction:

Article Title: Avian Tembusu virus infection effectively triggers host innate immune response through MDA5 and TLR3-dependent signaling pathways
Article Snippet: .. Equal amounts of RNA (4 μg) was used for reverse-transcription PCR to prepare cDNA using M-MLV Reverse Transcriptase (Promega, USA), followed by PCR using rTaq DNA polymerase and quantitative real-time PCR using TransStart Green qPCR SuperMix (TransGen). .. The primers for ATMUV and chicken IFN-β, IFN-λ gene were designed using the Primer 5 software; other sequences of the primers used in this study have been described previously [ – ].

Article Title: Cilia-related protein SPEF2 regulates osteoblast differentiation
Article Snippet: .. Real-time PCR (RT-PCR) For analysis of gene expression with RT-PCR the total RNA was reverse transcribed with random primers and an RT-PCR kit (ImProm-II Reverse Transcription System, Promega) according to the manufacturer’s instructions. .. Produced cDNA was amplified by using gene specific primers listed in Supplemental Table .

Reverse Transcription Polymerase Chain Reaction:

Article Title: Aspergillus fumigatus Survival in Alkaline and Extreme Zinc-Limiting Environments Relies on the Induction of a Zinc Homeostasis System Encoded by the zrfC and aspf2 Genes ▿ Genes ▿ †
Article Snippet: .. The cDNA of zrfC was obtained by reverse transcription (RT)-PCR using oligonucleotides JA299 (for retrotranscription) and JA60 and JA54 (both for PCR), and a 1.6-kb cDNA fragment was subcloned into pGEM-T Easy (Promega) to generate plasmid pZRF30 and sequenced. .. The cDNA of aspf2 was obtained by RT-PCR using oligonucleotides JA8 (for retrotranscription) and JA166 and JA167 (both for PCR), and a 0.96-kb cDNA fragment was subcloned into pGEM-T Easy to generate plasmid pASPF23 and sequenced.

Article Title: Restricted chromosomal silencing in nucleolar dominance
Article Snippet: .. Reverse transcription (RT)-PCR was performed by using RNA that had been treated with RQ1 DNase I (Promega) to eliminate any contaminating genomic DNA. .. Total RNA was used in all cases except in the case of F16N15.13 , for which poly(A)+ RNA was isolated on oligo(dT) paramagnetic beads according to the manufacturer's instructions (Dynal, Great Neck, NY).

Article Title: Effects of THAP11 on Erythroid Differentiation and Megakaryocytic Differentiation of K562 Cells
Article Snippet: .. Reverse transcription-PCR (RT-PCR) and quantitative real-time RT-PCR Total RNA was reverse-transcribed and amplified using reverse transcription and PCR kits, respectively (Promega Corp., Madison, WI, U.S.). .. Real-time RT-PCR was performed by Bio-Rad IQ5 (Bio-Rad, U.S.).

Article Title: Analysis of Transcription of the Staphylococcus aureus Aerobic Class Ib and Anaerobic Class III Ribonucleotide Reductase Genes in Response to Oxygen
Article Snippet: .. For reverse transcription (RT)-PCR and primer extension, residual DNA was removed by treatment with RQ1 RNase-free DNase (Promega). .. RNA concentrations were determined by A 260 measurements, and RNA integrity was analyzed by agarose/formaldehyde gel electrophoresis ( ).

Article Title: Virucidal nano-perforator of viral membrane trapping viral RNAs in the endosome
Article Snippet: .. To confirm the release of vRNPs, reverse transcription (RT)-PCR was carried out using M-MLV reverse transcriptase (M1705, Promega, Madison, WI, USA) according to the manufacturer’s protocol using primers for the viral M gene of A/Puerto Rico/8/34; 5′-TGCACTTTGACATTGTGGATTCTTG-3′ (M_FW) and 5′-CCCTCATAGACTTTGGCACTCC-3′ (M_BW) . .. The coding region of the M sequence in the RT product was amplified with the described primers by using rTaq Plus 5 × PCR Master Mix (ELPIS-BIOTECH, Inc., Daejeon, Korea) according to the manufacturer’s protocol under the following thermal cycling conditions: 30 cycles of 95 °C for 10 s, 56 °C for 10 s, and 72 °C for 10 s. PCR products were separated by electrophoresis at 135 V for 20 min on a 1% agarose (Cosmogenetech, Inc., Korea) gel and visualised with a UV transilluminator.

Article Title: Cilia-related protein SPEF2 regulates osteoblast differentiation
Article Snippet: .. Real-time PCR (RT-PCR) For analysis of gene expression with RT-PCR the total RNA was reverse transcribed with random primers and an RT-PCR kit (ImProm-II Reverse Transcription System, Promega) according to the manufacturer’s instructions. .. Produced cDNA was amplified by using gene specific primers listed in Supplemental Table .

Expressing:

Article Title: Cilia-related protein SPEF2 regulates osteoblast differentiation
Article Snippet: .. Real-time PCR (RT-PCR) For analysis of gene expression with RT-PCR the total RNA was reverse transcribed with random primers and an RT-PCR kit (ImProm-II Reverse Transcription System, Promega) according to the manufacturer’s instructions. .. Produced cDNA was amplified by using gene specific primers listed in Supplemental Table .

Polymerase Chain Reaction:

Article Title: Aspergillus fumigatus Survival in Alkaline and Extreme Zinc-Limiting Environments Relies on the Induction of a Zinc Homeostasis System Encoded by the zrfC and aspf2 Genes ▿ Genes ▿ †
Article Snippet: .. The cDNA of zrfC was obtained by reverse transcription (RT)-PCR using oligonucleotides JA299 (for retrotranscription) and JA60 and JA54 (both for PCR), and a 1.6-kb cDNA fragment was subcloned into pGEM-T Easy (Promega) to generate plasmid pZRF30 and sequenced. .. The cDNA of aspf2 was obtained by RT-PCR using oligonucleotides JA8 (for retrotranscription) and JA166 and JA167 (both for PCR), and a 0.96-kb cDNA fragment was subcloned into pGEM-T Easy to generate plasmid pASPF23 and sequenced.

Article Title: Avian Tembusu virus infection effectively triggers host innate immune response through MDA5 and TLR3-dependent signaling pathways
Article Snippet: .. Equal amounts of RNA (4 μg) was used for reverse-transcription PCR to prepare cDNA using M-MLV Reverse Transcriptase (Promega, USA), followed by PCR using rTaq DNA polymerase and quantitative real-time PCR using TransStart Green qPCR SuperMix (TransGen). .. The primers for ATMUV and chicken IFN-β, IFN-λ gene were designed using the Primer 5 software; other sequences of the primers used in this study have been described previously [ – ].

Article Title: Effects of THAP11 on Erythroid Differentiation and Megakaryocytic Differentiation of K562 Cells
Article Snippet: .. Reverse transcription-PCR (RT-PCR) and quantitative real-time RT-PCR Total RNA was reverse-transcribed and amplified using reverse transcription and PCR kits, respectively (Promega Corp., Madison, WI, U.S.). .. Real-time RT-PCR was performed by Bio-Rad IQ5 (Bio-Rad, U.S.).

Plasmid Preparation:

Article Title: Aspergillus fumigatus Survival in Alkaline and Extreme Zinc-Limiting Environments Relies on the Induction of a Zinc Homeostasis System Encoded by the zrfC and aspf2 Genes ▿ Genes ▿ †
Article Snippet: .. The cDNA of zrfC was obtained by reverse transcription (RT)-PCR using oligonucleotides JA299 (for retrotranscription) and JA60 and JA54 (both for PCR), and a 1.6-kb cDNA fragment was subcloned into pGEM-T Easy (Promega) to generate plasmid pZRF30 and sequenced. .. The cDNA of aspf2 was obtained by RT-PCR using oligonucleotides JA8 (for retrotranscription) and JA166 and JA167 (both for PCR), and a 0.96-kb cDNA fragment was subcloned into pGEM-T Easy to generate plasmid pASPF23 and sequenced.

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    Promega oligo dt primer
    USP21 modulates Hh signaling and Gli1 transcriptional activity. (A) NIH3T3 cells were treated with USP21-targeting siRNA oligonucleotides (#10, #11, #12) or a non-targeting control (NT3), serum-starved and then incubated with SAG (100 nM) for 4 h before mRNA extraction. USP21 and Gli1 mRNA levels were measured by performing quantitative reverse-transcription PCR and normalised to <t>RNA</t> levels of Pol2 ( n =3 independent experiments, mean±s.d.). x -axis labels given in bottom panel also apply to the top panel. (B) HEK293T cells were treated with USP21-targeting siRNA oligonucleotides (#5, #6) or non-targeting control <t>oligo</t> (NT3), or were treated with transfection reagent alone (Mock). 24 h before lysis, cells were transfected with Gli-responsive firefly luciferase and Renilla luciferase reporters (pGLB3B-12Gli-Luc and pRL-Renilla-TK), as well as with a Gli1 expression construct (HA–Gli1). Efficiency of the knockdown was assessed by western blotting ( n =2, bars show the range). (C) HEK293T cells were transfected with Gli-responsive firefly luciferase, Renilla luciferase reporters, HA–Gli1 and the indicated amounts of GFP, GFP–USP21 or GFP–USP21CS plasmids. Protein expression was assessed by western blotting (IB, immunoblot) for GFP. (GFP, n =5; USP21 and USP21CS, n =6; mean±s.d.; one-way ANOVA and Dunnett's test, ** P
    Oligo Dt Primer, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 317 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/oligo dt primer/product/Promega
    Average 99 stars, based on 317 article reviews
    Price from $9.99 to $1999.99
    oligo dt primer - by Bioz Stars, 2020-08
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    92
    Promega oligo dt chromatography
    : localisation of <t>mRNA.</t> (A) Fluorescent in situ hybridisation assay using specific <t>oligo</t> (dT) probe. As the control, RNase A was incubated with the parasites before probe hybridisation. DAPI: DNA stained with DAPI; mRNA: mRNA detection; granules: labelling of pixels with high intensity fluorescence signals; N: nucleus; K: kinetoplast. Scale bar = 5 µm. (B) Histogram of mRNA granule counts per cell.
    Oligo Dt Chromatography, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/oligo dt chromatography/product/Promega
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    oligo dt chromatography - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

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    USP21 modulates Hh signaling and Gli1 transcriptional activity. (A) NIH3T3 cells were treated with USP21-targeting siRNA oligonucleotides (#10, #11, #12) or a non-targeting control (NT3), serum-starved and then incubated with SAG (100 nM) for 4 h before mRNA extraction. USP21 and Gli1 mRNA levels were measured by performing quantitative reverse-transcription PCR and normalised to RNA levels of Pol2 ( n =3 independent experiments, mean±s.d.). x -axis labels given in bottom panel also apply to the top panel. (B) HEK293T cells were treated with USP21-targeting siRNA oligonucleotides (#5, #6) or non-targeting control oligo (NT3), or were treated with transfection reagent alone (Mock). 24 h before lysis, cells were transfected with Gli-responsive firefly luciferase and Renilla luciferase reporters (pGLB3B-12Gli-Luc and pRL-Renilla-TK), as well as with a Gli1 expression construct (HA–Gli1). Efficiency of the knockdown was assessed by western blotting ( n =2, bars show the range). (C) HEK293T cells were transfected with Gli-responsive firefly luciferase, Renilla luciferase reporters, HA–Gli1 and the indicated amounts of GFP, GFP–USP21 or GFP–USP21CS plasmids. Protein expression was assessed by western blotting (IB, immunoblot) for GFP. (GFP, n =5; USP21 and USP21CS, n =6; mean±s.d.; one-way ANOVA and Dunnett's test, ** P

    Journal: Journal of Cell Science

    Article Title: The centrosomal deubiquitylase USP21 regulates Gli1 transcriptional activity and stability

    doi: 10.1242/jcs.188516

    Figure Lengend Snippet: USP21 modulates Hh signaling and Gli1 transcriptional activity. (A) NIH3T3 cells were treated with USP21-targeting siRNA oligonucleotides (#10, #11, #12) or a non-targeting control (NT3), serum-starved and then incubated with SAG (100 nM) for 4 h before mRNA extraction. USP21 and Gli1 mRNA levels were measured by performing quantitative reverse-transcription PCR and normalised to RNA levels of Pol2 ( n =3 independent experiments, mean±s.d.). x -axis labels given in bottom panel also apply to the top panel. (B) HEK293T cells were treated with USP21-targeting siRNA oligonucleotides (#5, #6) or non-targeting control oligo (NT3), or were treated with transfection reagent alone (Mock). 24 h before lysis, cells were transfected with Gli-responsive firefly luciferase and Renilla luciferase reporters (pGLB3B-12Gli-Luc and pRL-Renilla-TK), as well as with a Gli1 expression construct (HA–Gli1). Efficiency of the knockdown was assessed by western blotting ( n =2, bars show the range). (C) HEK293T cells were transfected with Gli-responsive firefly luciferase, Renilla luciferase reporters, HA–Gli1 and the indicated amounts of GFP, GFP–USP21 or GFP–USP21CS plasmids. Protein expression was assessed by western blotting (IB, immunoblot) for GFP. (GFP, n =5; USP21 and USP21CS, n =6; mean±s.d.; one-way ANOVA and Dunnett's test, ** P

    Article Snippet: Quantitative reverse-transcription PCR Cells were lysed, and mRNA was extracted using the RNAeasy mini kit (Qiagen). cDNA synthesis was performed using 1 µg RNA with RevertAid H-minus M-MuLV reverse transcriptase (Fermentas) using an oligo-dT primer (Promega).

    Techniques: Activity Assay, Incubation, Polymerase Chain Reaction, Transfection, Lysis, Luciferase, Expressing, Construct, Western Blot

    FcRn expression in normal adult human small intestine and human intestinal epithelial cell lines. Western blots of total cellular protein (13 μg protein per lane, a ; 10 μg protein per lane, b ) isolated from the indicated source using affinity-purified rabbit antisera raised against amino acids 112–125 ( a ) or amino acids 174–188 ( b ). ( c ) RT-PCR detection of FcγRI transcripts. Total RNA (2 μg) from T84 (lanes 3 and 4), MOLT-4 (lanes 5 and 6; negative control), and U937 (lanes 1 and 2; positive control) cell lines was incubated with an oligo-dT primer with (odd-numbered lanes) or without (even-numbered lanes) avian myeloblastosis virus–RT (AMV-RT), and a nested PCR was performed with primers specific for FcγRI cDNA (top) or for β-actin (bottom).

    Journal: Journal of Clinical Investigation

    Article Title: Bidirectional FcRn-dependent IgG transport in a polarized human intestinal epithelial cell line

    doi:

    Figure Lengend Snippet: FcRn expression in normal adult human small intestine and human intestinal epithelial cell lines. Western blots of total cellular protein (13 μg protein per lane, a ; 10 μg protein per lane, b ) isolated from the indicated source using affinity-purified rabbit antisera raised against amino acids 112–125 ( a ) or amino acids 174–188 ( b ). ( c ) RT-PCR detection of FcγRI transcripts. Total RNA (2 μg) from T84 (lanes 3 and 4), MOLT-4 (lanes 5 and 6; negative control), and U937 (lanes 1 and 2; positive control) cell lines was incubated with an oligo-dT primer with (odd-numbered lanes) or without (even-numbered lanes) avian myeloblastosis virus–RT (AMV-RT), and a nested PCR was performed with primers specific for FcγRI cDNA (top) or for β-actin (bottom).

    Article Snippet: RNA (2 μg) was reverse-transcribed to cDNA with an oligo-dT primer (Promega Corp., Madison, Wisconsin, USA) and avian myeloblastosis virus reverse transcriptase (Promega Corp.).

    Techniques: Expressing, Western Blot, Isolation, Affinity Purification, Reverse Transcription Polymerase Chain Reaction, Negative Control, Positive Control, Incubation, Nested PCR

    A. Genomic location of ORF 20 and ORF 50 relative to adjacent genes. Genes are indicated by open boxes, arrow head represents direction of transcription. Dotted lines represent introns. The nucleotide positions representing the location of the predicted TATA box and poly A site for each gene are indicated. The position of primers used for PCR and cDNA priming (ORF 20) are indicated by arrows and nucleotide position. Predicted miRNA binding sequences are indicated by vertical bars in the respective UTRs. B. ORF 20 Lane 1, No RT; Lane 2, cDNA primed with 35 pm primer (250 ng); Lane 3, cDNA primed with 10 pm primer (66ng); Lane 4,: No template control; Lane 5, DNA positive control. Lane 6; Marker, Generuler 100bp C. ORF 50 Lane 1, No RT, Lane 2, cDNA primed with Oligo dT; Lane 3, cDNA primed with random primers; Lane 4, No template control; Lane 5, DNA positive control; Lane 6, Marker, Generuler 1kb D. ORF73 Lane 1, No RT, Lane 2, cDNA primed with Oligo dT; Lane 3, cDNA primed with random primers; Lane 4, No template control; Lane 5, DNA positive control; Lane 6, Marker, Generuler 100bp

    Journal: The Journal of general virology

    Article Title: Ovine herpesvirus-2 encoded microRNAs target virus genes involved in virus latency

    doi: 10.1099/vir.0.059303-0

    Figure Lengend Snippet: A. Genomic location of ORF 20 and ORF 50 relative to adjacent genes. Genes are indicated by open boxes, arrow head represents direction of transcription. Dotted lines represent introns. The nucleotide positions representing the location of the predicted TATA box and poly A site for each gene are indicated. The position of primers used for PCR and cDNA priming (ORF 20) are indicated by arrows and nucleotide position. Predicted miRNA binding sequences are indicated by vertical bars in the respective UTRs. B. ORF 20 Lane 1, No RT; Lane 2, cDNA primed with 35 pm primer (250 ng); Lane 3, cDNA primed with 10 pm primer (66ng); Lane 4,: No template control; Lane 5, DNA positive control. Lane 6; Marker, Generuler 100bp C. ORF 50 Lane 1, No RT, Lane 2, cDNA primed with Oligo dT; Lane 3, cDNA primed with random primers; Lane 4, No template control; Lane 5, DNA positive control; Lane 6, Marker, Generuler 1kb D. ORF73 Lane 1, No RT, Lane 2, cDNA primed with Oligo dT; Lane 3, cDNA primed with random primers; Lane 4, No template control; Lane 5, DNA positive control; Lane 6, Marker, Generuler 100bp

    Article Snippet: 1 μg of RNA was digested with RQ1 Dnase (Promega) for 30 min at 37 °C. cDNA was primed using 250ng Oligo dT primer (Promega; equivalent to 0.5 μg/μg RNA) and synthesized using AMV reverse transcriptase for 1 hr at 42 °C.

    Techniques: Polymerase Chain Reaction, Binding Assay, Positive Control, Marker

    : localisation of mRNA. (A) Fluorescent in situ hybridisation assay using specific oligo (dT) probe. As the control, RNase A was incubated with the parasites before probe hybridisation. DAPI: DNA stained with DAPI; mRNA: mRNA detection; granules: labelling of pixels with high intensity fluorescence signals; N: nucleus; K: kinetoplast. Scale bar = 5 µm. (B) Histogram of mRNA granule counts per cell.

    Journal: Memórias do Instituto Oswaldo Cruz

    Article Title: Trypanosoma cruzi transcriptome during axenic epimastigote growth curve

    doi: 10.1590/0074-02760170404

    Figure Lengend Snippet: : localisation of mRNA. (A) Fluorescent in situ hybridisation assay using specific oligo (dT) probe. As the control, RNase A was incubated with the parasites before probe hybridisation. DAPI: DNA stained with DAPI; mRNA: mRNA detection; granules: labelling of pixels with high intensity fluorescence signals; N: nucleus; K: kinetoplast. Scale bar = 5 µm. (B) Histogram of mRNA granule counts per cell.

    Article Snippet: Subsequently, RNA integrity and quality was analysed using the Agilent 2100 bioanalyzer and the RNA 6000 Pico Kit (Agilent). mRNA was selected by oligo-dT chromatography using the PolyATtract® mRNA Isolation Kit (Promega).

    Techniques: In Situ, Hybridization, Incubation, Staining, Fluorescence