superscript iii first strand synthesis supermix kit  (Thermo Fisher)


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    Name:
    SuperScript III First Strand Synthesis SuperMix for qRT PCR
    Description:
    SuperScript III First Strand Synthesis SuperMix for qRT PCR provides the high temperature capability of SuperScript III Reverse Transcriptase in an optimized SuperMix format for the synthesis of first strand cDNA for use in real time quantitative RT PCR qRT PCR The simple time saving reaction set up uses just two tubes a 2X Reaction Mix and an Enzyme Mix Enzyme mixSuperScript III Reverse Transcriptase included in the RT Enzyme Mix is a version of M MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability The enzyme can be used to synthesize cDNA at a temperature range of 42 60°C providing increased specificity higher yields of cDNA and more full length product than other reverse transcriptases Because SuperScript III RT is not significantly inhibited by ribosomal and transfer RNA it can be used to synthesize cDNA from total RNA RNaseOUT Recombinant Ribonuclease Inhibitor also included in the enzyme mix is an RNase inhibitor protein that safeguards against the degradation of target RNA due to ribonuclease contamination of the RNA preparation Reaction mixThe 2X RT Reaction Mix includes oligo dT 20 random hexamers MgCl2 and dNTPs in a buffer formulation that has been optimized for qRT PCR E coli RNase H is provided as a separate tube in the kit to remove the RNA template from the cDNA RNA hybrid molecule after first strand synthesis This has been shown to increase sensitivity in qRT PCR Using SuperScript III First Strand Synthesis SuperMixThis SuperMix formulation can be used to quantify fewer than 10 copies of a target gene in qRT PCR with a broad dynamic range that supports accurate quantification of high copy mRNA from up to 1 µg of total RNA Reagents are provided for 50 or 250 RT reactions of 20 µL each
    Catalog Number:
    11752050
    Price:
    None
    Category:
    Kits and Assays
    Applications:
    PCR & Real-Time PCR|Reverse Transcription
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    Structured Review

    Thermo Fisher superscript iii first strand synthesis supermix kit
    SuperScript III First Strand Synthesis SuperMix for qRT PCR provides the high temperature capability of SuperScript III Reverse Transcriptase in an optimized SuperMix format for the synthesis of first strand cDNA for use in real time quantitative RT PCR qRT PCR The simple time saving reaction set up uses just two tubes a 2X Reaction Mix and an Enzyme Mix Enzyme mixSuperScript III Reverse Transcriptase included in the RT Enzyme Mix is a version of M MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability The enzyme can be used to synthesize cDNA at a temperature range of 42 60°C providing increased specificity higher yields of cDNA and more full length product than other reverse transcriptases Because SuperScript III RT is not significantly inhibited by ribosomal and transfer RNA it can be used to synthesize cDNA from total RNA RNaseOUT Recombinant Ribonuclease Inhibitor also included in the enzyme mix is an RNase inhibitor protein that safeguards against the degradation of target RNA due to ribonuclease contamination of the RNA preparation Reaction mixThe 2X RT Reaction Mix includes oligo dT 20 random hexamers MgCl2 and dNTPs in a buffer formulation that has been optimized for qRT PCR E coli RNase H is provided as a separate tube in the kit to remove the RNA template from the cDNA RNA hybrid molecule after first strand synthesis This has been shown to increase sensitivity in qRT PCR Using SuperScript III First Strand Synthesis SuperMixThis SuperMix formulation can be used to quantify fewer than 10 copies of a target gene in qRT PCR with a broad dynamic range that supports accurate quantification of high copy mRNA from up to 1 µg of total RNA Reagents are provided for 50 or 250 RT reactions of 20 µL each
    https://www.bioz.com/result/superscript iii first strand synthesis supermix kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    superscript iii first strand synthesis supermix kit - by Bioz Stars, 2021-03
    99/100 stars

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    Related Articles

    Quantitative RT-PCR:

    Article Title: The MT1G Gene in LUHMES Neurons Is a Sensitive Biomarker of Neurotoxicity
    Article Snippet: .. qRT-PCR For gene expression assays, 3 × 105 cells were cultured per well in 12 well plates with 2 mL medium to enable RNA extraction and quantitative reverse transcriptase PCR (qRT-PCR). .. Cells were treated with each toxicant for 6 h in 2 mL of medium in 12-well microplates.

    Article Title: Flow-cytometric microglial sorting coupled with quantitative proteomics identifies moesin as a highly-abundant microglial protein with relevance to Alzheimer’s disease
    Article Snippet: Small interfering RNA (siRNA) transfection studiesPrimary microglia were transfected with 40nM (final concentration) of either Msn siRNA (Santa Cruz Biotechnology, Cat. No. sc-35956) or nonspecific sham siRNA (Santa Cruz Biotechnology, Cat. No. sc-37007) using Lipofectamine™ RNAiMAX (Invitrogen, Cat. No. 13778100) and Opti-MEM (Invitrogen, Cat. No. 31985-062). .. After 48 hours, the efficiency of siRNA-mediated gene silencing was confirmed by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) ( ). .. Quantitative reverse transcriptase PCR (qRT‐PCR)Total RNA from microglia was extracted using Trizol (Invitrogen, Cat. No. 15596026) and an RNeasy mini extraction kit (Qiagen, Cat. No. 74104) according to the manufacturer’s instructions.

    Article Title: Increased Bacterial Load and Expression of Antimicrobial Peptides in Skin of Barrier-Deficient Mice with Reduced Cancer Susceptibility
    Article Snippet: The RNeasy mini kit (Qiagen, West Sussex, UK) was used according to the manufacturer’s instructions. .. RNA concentration was measured using an ND-100 NanoDrop spectrophotometer (NanoDrop Technologies, Wilmington, DE). cDNA was synthesized using a Superscript III First-Strand Synthesis Supermix for quantitative reverse transcriptase PCR (qRT-PCR) kit (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. qRT-PCR was carried out using specific primers and fast SYBR Green and run on an ABI Prism 7900HT Sequence detection system. ..

    Article Title: Viral Determinants in H5N1 Influenza A Virus Enable Productive Infection of HeLa Cells
    Article Snippet: At 48 h posttransfection, either cells were infected, and productive virus growth was measured by plaque assay, or cell viability was determined using the CellTiter-Glo assay (Promega). .. RNA extraction and quantitative reverse transcription-PCR (qRT-PCR). .. At 48 h post-siRNA transfection, total RNA was extracted from HeLa cells using an RNeasy minikit (Qiagen).

    Article Title: Production and Functional Characterization of a Recombinant Predicted Pore-Forming Protein (TVSAPLIP12) of Trichomonas vaginalis in Nicotiana benthamiana Plants
    Article Snippet: Complementary DNA (cDNA) was obtained from 1 μg of total RNA using SuperScript IV Reverse Transcriptase and Random Hexamers (Invitrogen) as primers, following manufacturer's instructions. .. Transcription of selected TvSaplip genes 1–8, 11, 12 (GenBank accession no: TVAG_388060 , TVAG_209200 , TVAG_393030 , TVAG_473630 , TVAG_000220 , TVAG_453350 , TVAG_070250 , TVAG_213250 , TVAG_306610 , TVAG_183780 ) was evaluated by quantitative Reverse Transcription PCR (qRT-PCR). .. Specific TaqMan primer/probe sets have been designed by Beacon Designer Software (Premier Biosoft) for all TvSaplip genes except TvSaplip7 .

    Expressing:

    Article Title: The MT1G Gene in LUHMES Neurons Is a Sensitive Biomarker of Neurotoxicity
    Article Snippet: .. qRT-PCR For gene expression assays, 3 × 105 cells were cultured per well in 12 well plates with 2 mL medium to enable RNA extraction and quantitative reverse transcriptase PCR (qRT-PCR). .. Cells were treated with each toxicant for 6 h in 2 mL of medium in 12-well microplates.

    Cell Culture:

    Article Title: The MT1G Gene in LUHMES Neurons Is a Sensitive Biomarker of Neurotoxicity
    Article Snippet: .. qRT-PCR For gene expression assays, 3 × 105 cells were cultured per well in 12 well plates with 2 mL medium to enable RNA extraction and quantitative reverse transcriptase PCR (qRT-PCR). .. Cells were treated with each toxicant for 6 h in 2 mL of medium in 12-well microplates.

    RNA Extraction:

    Article Title: The MT1G Gene in LUHMES Neurons Is a Sensitive Biomarker of Neurotoxicity
    Article Snippet: .. qRT-PCR For gene expression assays, 3 × 105 cells were cultured per well in 12 well plates with 2 mL medium to enable RNA extraction and quantitative reverse transcriptase PCR (qRT-PCR). .. Cells were treated with each toxicant for 6 h in 2 mL of medium in 12-well microplates.

    Article Title: Viral Determinants in H5N1 Influenza A Virus Enable Productive Infection of HeLa Cells
    Article Snippet: At 48 h posttransfection, either cells were infected, and productive virus growth was measured by plaque assay, or cell viability was determined using the CellTiter-Glo assay (Promega). .. RNA extraction and quantitative reverse transcription-PCR (qRT-PCR). .. At 48 h post-siRNA transfection, total RNA was extracted from HeLa cells using an RNeasy minikit (Qiagen).

    Polymerase Chain Reaction:

    Article Title: The MT1G Gene in LUHMES Neurons Is a Sensitive Biomarker of Neurotoxicity
    Article Snippet: .. qRT-PCR For gene expression assays, 3 × 105 cells were cultured per well in 12 well plates with 2 mL medium to enable RNA extraction and quantitative reverse transcriptase PCR (qRT-PCR). .. Cells were treated with each toxicant for 6 h in 2 mL of medium in 12-well microplates.

    Article Title: Flow-cytometric microglial sorting coupled with quantitative proteomics identifies moesin as a highly-abundant microglial protein with relevance to Alzheimer’s disease
    Article Snippet: Small interfering RNA (siRNA) transfection studiesPrimary microglia were transfected with 40nM (final concentration) of either Msn siRNA (Santa Cruz Biotechnology, Cat. No. sc-35956) or nonspecific sham siRNA (Santa Cruz Biotechnology, Cat. No. sc-37007) using Lipofectamine™ RNAiMAX (Invitrogen, Cat. No. 13778100) and Opti-MEM (Invitrogen, Cat. No. 31985-062). .. After 48 hours, the efficiency of siRNA-mediated gene silencing was confirmed by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) ( ). .. Quantitative reverse transcriptase PCR (qRT‐PCR)Total RNA from microglia was extracted using Trizol (Invitrogen, Cat. No. 15596026) and an RNeasy mini extraction kit (Qiagen, Cat. No. 74104) according to the manufacturer’s instructions.

    Article Title: Increased Bacterial Load and Expression of Antimicrobial Peptides in Skin of Barrier-Deficient Mice with Reduced Cancer Susceptibility
    Article Snippet: The RNeasy mini kit (Qiagen, West Sussex, UK) was used according to the manufacturer’s instructions. .. RNA concentration was measured using an ND-100 NanoDrop spectrophotometer (NanoDrop Technologies, Wilmington, DE). cDNA was synthesized using a Superscript III First-Strand Synthesis Supermix for quantitative reverse transcriptase PCR (qRT-PCR) kit (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. qRT-PCR was carried out using specific primers and fast SYBR Green and run on an ABI Prism 7900HT Sequence detection system. ..

    Article Title: Production and Functional Characterization of a Recombinant Predicted Pore-Forming Protein (TVSAPLIP12) of Trichomonas vaginalis in Nicotiana benthamiana Plants
    Article Snippet: Complementary DNA (cDNA) was obtained from 1 μg of total RNA using SuperScript IV Reverse Transcriptase and Random Hexamers (Invitrogen) as primers, following manufacturer's instructions. .. Transcription of selected TvSaplip genes 1–8, 11, 12 (GenBank accession no: TVAG_388060 , TVAG_209200 , TVAG_393030 , TVAG_473630 , TVAG_000220 , TVAG_453350 , TVAG_070250 , TVAG_213250 , TVAG_306610 , TVAG_183780 ) was evaluated by quantitative Reverse Transcription PCR (qRT-PCR). .. Specific TaqMan primer/probe sets have been designed by Beacon Designer Software (Premier Biosoft) for all TvSaplip genes except TvSaplip7 .

    Article Title: HIV-1 uncoating by release of viral cDNA from capsid-like structures in the nucleus of infected cells
    Article Snippet: 8 h prior to RNA extraction and purification using the Invitrap spin universal RNA mini kit (Stratec biomedical, Germany), medium was changed for medium containing 1-25 μM flavopiridol (Sigma Aldrich) or 1-25 μM 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB; Sigma Aldrich) as indicated. .. Quantitative reverse transcription PCR was performed as previously described ( ). .. Briefly, messenger RNA (mRNA) levels were quantified by TaqMan quantitative RT-PCR (qRT-PCR).

    Concentration Assay:

    Article Title: Increased Bacterial Load and Expression of Antimicrobial Peptides in Skin of Barrier-Deficient Mice with Reduced Cancer Susceptibility
    Article Snippet: The RNeasy mini kit (Qiagen, West Sussex, UK) was used according to the manufacturer’s instructions. .. RNA concentration was measured using an ND-100 NanoDrop spectrophotometer (NanoDrop Technologies, Wilmington, DE). cDNA was synthesized using a Superscript III First-Strand Synthesis Supermix for quantitative reverse transcriptase PCR (qRT-PCR) kit (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. qRT-PCR was carried out using specific primers and fast SYBR Green and run on an ABI Prism 7900HT Sequence detection system. ..

    Spectrophotometry:

    Article Title: Increased Bacterial Load and Expression of Antimicrobial Peptides in Skin of Barrier-Deficient Mice with Reduced Cancer Susceptibility
    Article Snippet: The RNeasy mini kit (Qiagen, West Sussex, UK) was used according to the manufacturer’s instructions. .. RNA concentration was measured using an ND-100 NanoDrop spectrophotometer (NanoDrop Technologies, Wilmington, DE). cDNA was synthesized using a Superscript III First-Strand Synthesis Supermix for quantitative reverse transcriptase PCR (qRT-PCR) kit (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. qRT-PCR was carried out using specific primers and fast SYBR Green and run on an ABI Prism 7900HT Sequence detection system. ..

    Synthesized:

    Article Title: Increased Bacterial Load and Expression of Antimicrobial Peptides in Skin of Barrier-Deficient Mice with Reduced Cancer Susceptibility
    Article Snippet: The RNeasy mini kit (Qiagen, West Sussex, UK) was used according to the manufacturer’s instructions. .. RNA concentration was measured using an ND-100 NanoDrop spectrophotometer (NanoDrop Technologies, Wilmington, DE). cDNA was synthesized using a Superscript III First-Strand Synthesis Supermix for quantitative reverse transcriptase PCR (qRT-PCR) kit (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. qRT-PCR was carried out using specific primers and fast SYBR Green and run on an ABI Prism 7900HT Sequence detection system. ..

    SYBR Green Assay:

    Article Title: Increased Bacterial Load and Expression of Antimicrobial Peptides in Skin of Barrier-Deficient Mice with Reduced Cancer Susceptibility
    Article Snippet: The RNeasy mini kit (Qiagen, West Sussex, UK) was used according to the manufacturer’s instructions. .. RNA concentration was measured using an ND-100 NanoDrop spectrophotometer (NanoDrop Technologies, Wilmington, DE). cDNA was synthesized using a Superscript III First-Strand Synthesis Supermix for quantitative reverse transcriptase PCR (qRT-PCR) kit (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. qRT-PCR was carried out using specific primers and fast SYBR Green and run on an ABI Prism 7900HT Sequence detection system. ..

    Sequencing:

    Article Title: Increased Bacterial Load and Expression of Antimicrobial Peptides in Skin of Barrier-Deficient Mice with Reduced Cancer Susceptibility
    Article Snippet: The RNeasy mini kit (Qiagen, West Sussex, UK) was used according to the manufacturer’s instructions. .. RNA concentration was measured using an ND-100 NanoDrop spectrophotometer (NanoDrop Technologies, Wilmington, DE). cDNA was synthesized using a Superscript III First-Strand Synthesis Supermix for quantitative reverse transcriptase PCR (qRT-PCR) kit (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. qRT-PCR was carried out using specific primers and fast SYBR Green and run on an ABI Prism 7900HT Sequence detection system. ..

    Isolation:

    Article Title: Rsp promotes the transcription of virulence factors in an agr-independent manner in Staphylococcus aureus
    Article Snippet: The combinant plasmids were first electroporated into S. aureus strain RN4220 for modification and subsequently transformed into the WT and agr mutant strains to derive the overexpressed strains. .. Total RNA isolation, cDNA generation, and real-time quantitative reverse transcription-PCR Overnight cultures of S. aureus were diluted 1:100 in TSB and then grown to the indicated cell density until being collected. .. S. aureus cells were collected by centrifugation and processed with 1 ml of RNAiso plus (TaKaRa) in combination with 0.1-mm-diameter silica-zirconia beads in a FastPrep-24 automated system (MP Biomedicals), and the residual DNA was removed with RNase-free DNase I (TransGen, 3 U/μl).

    Article Title: Latrunculin A-Induced Perturbation of the Actin Cytoskeleton Mediates Pap1p-Dependent Induction of the Caf5p Efflux Pump in Schizosaccharomyces pombe
    Article Snippet: S. pombe cells expressing Pap1-GFP fusions were observed live (GFP filter set) using a Leica DMI6000B inverted microscope equipped with a 100 × Plan Apochromat 1.4 NA oil objective and a Photometrics QuantEM:512SC EMCCD camera driven by Metamorph software. .. qRT-PCR Total RNA was isolated using TRIzol Reagent (Life Technologies) according to the supplier’s protocol. cDNA synthesis was performed with 1 µg of total RNA using the SuperScript III First Strand Synthesis Supermix for qRT-PCR kit (Invitrogen), according to the supplier’s protocol. .. Real-time PCR was performed using a Bio-Rad CFX Connect Real-Time PCR Detection System in conjunction with the Maxima SYBR Green qPCR Master Mix kit (Thermo Scientific).

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    • reverse transcriptase pcr qrt pcr kit  (Thermo Fisher)
    99
    Thermo Fisher reverse transcriptase pcr qrt pcr kit
    Microbiota independent inflammatory and stress response phenotypes of EPI-/- skin. ( a ) qPCR of 16s RNA gene in WT, EPI-/-, antibiotic-treated EPI-/-, and flora-deficient EPI-/- ear skin samples. ( b ) Number of CD4 + CD3 + cells per mm 2 dermis. ( c ) Number of γδTCR + cells per mm epidermis. ( d ) Number of Vγ3 + cells per mm epidermis. ( e ) <t>qRT-PCR</t> of Rae-1 in WT, EPI-/-, antibiotic-treated EPI-/-, and flora-deficient EPI-/- epidermis. ( f ) Quantification of MPO in WT, EPI-/-, and flora-deficient skin lysates. Data are means ± SEM from at least four ( a ) or three ( b–f ) mice per group. * P
    Reverse Transcriptase Pcr Qrt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/reverse transcriptase pcr qrt pcr kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    reverse transcriptase pcr qrt pcr kit - by Bioz Stars, 2021-03
    99/100 stars
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    Microbiota independent inflammatory and stress response phenotypes of EPI-/- skin. ( a ) qPCR of 16s RNA gene in WT, EPI-/-, antibiotic-treated EPI-/-, and flora-deficient EPI-/- ear skin samples. ( b ) Number of CD4 + CD3 + cells per mm 2 dermis. ( c ) Number of γδTCR + cells per mm epidermis. ( d ) Number of Vγ3 + cells per mm epidermis. ( e ) qRT-PCR of Rae-1 in WT, EPI-/-, antibiotic-treated EPI-/-, and flora-deficient EPI-/- epidermis. ( f ) Quantification of MPO in WT, EPI-/-, and flora-deficient skin lysates. Data are means ± SEM from at least four ( a ) or three ( b–f ) mice per group. * P

    Journal: The Journal of Investigative Dermatology

    Article Title: Increased Bacterial Load and Expression of Antimicrobial Peptides in Skin of Barrier-Deficient Mice with Reduced Cancer Susceptibility

    doi: 10.1038/JID.2015.383

    Figure Lengend Snippet: Microbiota independent inflammatory and stress response phenotypes of EPI-/- skin. ( a ) qPCR of 16s RNA gene in WT, EPI-/-, antibiotic-treated EPI-/-, and flora-deficient EPI-/- ear skin samples. ( b ) Number of CD4 + CD3 + cells per mm 2 dermis. ( c ) Number of γδTCR + cells per mm epidermis. ( d ) Number of Vγ3 + cells per mm epidermis. ( e ) qRT-PCR of Rae-1 in WT, EPI-/-, antibiotic-treated EPI-/-, and flora-deficient EPI-/- epidermis. ( f ) Quantification of MPO in WT, EPI-/-, and flora-deficient skin lysates. Data are means ± SEM from at least four ( a ) or three ( b–f ) mice per group. * P

    Article Snippet: RNA concentration was measured using an ND-100 NanoDrop spectrophotometer (NanoDrop Technologies, Wilmington, DE). cDNA was synthesized using a Superscript III First-Strand Synthesis Supermix for quantitative reverse transcriptase PCR (qRT-PCR) kit (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. qRT-PCR was carried out using specific primers and fast SYBR Green and run on an ABI Prism 7900HT Sequence detection system.

    Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Mouse Assay

    AMP expression and protease activity in EPI-/- epidermis after microbiota modulation. ( a–k ) qRT-PCR of Camp ( a ) , Defb1 ( b ), Defb2 ( c ), Defb3 ( d ), Defb6 ( e ), S100A9 ( f ), Pla2g2a ( g ), Reg3b ( h ), Reg3g ( i ), and Serpina1b ( j ) in WT, EPI-/-, antibiotic-treated EPI-/-, and flora-deficient EPI-/- epidermis. Data are means ± SEM from at least four mice per group. * P

    Journal: The Journal of Investigative Dermatology

    Article Title: Increased Bacterial Load and Expression of Antimicrobial Peptides in Skin of Barrier-Deficient Mice with Reduced Cancer Susceptibility

    doi: 10.1038/JID.2015.383

    Figure Lengend Snippet: AMP expression and protease activity in EPI-/- epidermis after microbiota modulation. ( a–k ) qRT-PCR of Camp ( a ) , Defb1 ( b ), Defb2 ( c ), Defb3 ( d ), Defb6 ( e ), S100A9 ( f ), Pla2g2a ( g ), Reg3b ( h ), Reg3g ( i ), and Serpina1b ( j ) in WT, EPI-/-, antibiotic-treated EPI-/-, and flora-deficient EPI-/- epidermis. Data are means ± SEM from at least four mice per group. * P

    Article Snippet: RNA concentration was measured using an ND-100 NanoDrop spectrophotometer (NanoDrop Technologies, Wilmington, DE). cDNA was synthesized using a Superscript III First-Strand Synthesis Supermix for quantitative reverse transcriptase PCR (qRT-PCR) kit (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. qRT-PCR was carried out using specific primers and fast SYBR Green and run on an ABI Prism 7900HT Sequence detection system.

    Techniques: Expressing, Activity Assay, Quantitative RT-PCR, Mouse Assay

    The caf5 gene is induced in a pap1 -dependent manner in response to LatA. qRT-PCR analysis of caf5 and trr1 expression (relative to gpd1 ) in response to 0.3 μM LatA treatment in (A) wild-type, (B) pap1 Δ, and (C) act1-R183A , D184A backgrounds. The level of caf5 and trr1 transcripts were normalized to that of the internal control gpd1 . The normalized level of the transcripts in LatA-treated cells is shown relative to that of DMSO controls. The mean ± SE for three biological replicates is shown. DMSO, dimethyl sulfoxide; LatA, latrunculin A; qRT-PCR, quantitative real time-polymerase chain reaction.

    Journal: G3: Genes|Genomes|Genetics

    Article Title: Latrunculin A-Induced Perturbation of the Actin Cytoskeleton Mediates Pap1p-Dependent Induction of the Caf5p Efflux Pump in Schizosaccharomyces pombe

    doi: 10.1534/g3.116.037903

    Figure Lengend Snippet: The caf5 gene is induced in a pap1 -dependent manner in response to LatA. qRT-PCR analysis of caf5 and trr1 expression (relative to gpd1 ) in response to 0.3 μM LatA treatment in (A) wild-type, (B) pap1 Δ, and (C) act1-R183A , D184A backgrounds. The level of caf5 and trr1 transcripts were normalized to that of the internal control gpd1 . The normalized level of the transcripts in LatA-treated cells is shown relative to that of DMSO controls. The mean ± SE for three biological replicates is shown. DMSO, dimethyl sulfoxide; LatA, latrunculin A; qRT-PCR, quantitative real time-polymerase chain reaction.

    Article Snippet: qRT-PCR Total RNA was isolated using TRIzol Reagent (Life Technologies) according to the supplier’s protocol. cDNA synthesis was performed with 1 µg of total RNA using the SuperScript III First Strand Synthesis Supermix for qRT-PCR kit (Invitrogen), according to the supplier’s protocol.

    Techniques: Quantitative RT-PCR, Expressing, Real-time Polymerase Chain Reaction

    L-WNT3A treated autografts exhibit enhanced survival and osteogenesis. ( A ) Experimental design, where bone grafts were harvested, separated into mineralized matrix and marrow components then treated with either L-PBS or L-WNT3A. ( B ) qRT-PCR analyses of both components, showing fold-change in expression of Axin2 , Runx2 , and Osterix 24 h after treatment with L-PBS (white bars) or L-WNT3A (grey bars). TUNEL staining on representative tissue sections derived from aliquots of the mineralized matrix component of a bone graft treated with either ( C ) L-PBS or ( D ) L-WNT3A for 1 h then transplanted to the SRC for 10 days. ( E ) Quantification of TUNEL +ve cells/total number of viable, DAPI +ve cells on representative tissue sections. ALP staining on representative tissue sections derived from aliquots of the mineralized matrix component of a bone graft treated with either ( F ) L-PBS or ( G ) L-WNT3A; ( H ) quantification of ALP +ve pixels/total pixels. Aniline blue staining on representative tissue sections derived from aliquots of the mineralized matrix component of a bone graft treated with either ( I ) L-PBS or ( J ) L-WNT3A; ( K ) quantification of new bone volume. Scale bars = 50 µm. Asterisk indicates p-value

    Journal: Scientific Reports

    Article Title: A WNT protein therapeutic improves the bone-forming capacity of autografts from aged animals

    doi: 10.1038/s41598-017-18375-x

    Figure Lengend Snippet: L-WNT3A treated autografts exhibit enhanced survival and osteogenesis. ( A ) Experimental design, where bone grafts were harvested, separated into mineralized matrix and marrow components then treated with either L-PBS or L-WNT3A. ( B ) qRT-PCR analyses of both components, showing fold-change in expression of Axin2 , Runx2 , and Osterix 24 h after treatment with L-PBS (white bars) or L-WNT3A (grey bars). TUNEL staining on representative tissue sections derived from aliquots of the mineralized matrix component of a bone graft treated with either ( C ) L-PBS or ( D ) L-WNT3A for 1 h then transplanted to the SRC for 10 days. ( E ) Quantification of TUNEL +ve cells/total number of viable, DAPI +ve cells on representative tissue sections. ALP staining on representative tissue sections derived from aliquots of the mineralized matrix component of a bone graft treated with either ( F ) L-PBS or ( G ) L-WNT3A; ( H ) quantification of ALP +ve pixels/total pixels. Aniline blue staining on representative tissue sections derived from aliquots of the mineralized matrix component of a bone graft treated with either ( I ) L-PBS or ( J ) L-WNT3A; ( K ) quantification of new bone volume. Scale bars = 50 µm. Asterisk indicates p-value

    Article Snippet: Reverse transcription was performed with SuperScript III First-Strand Synthesis SuperMix for qRT-PCR Kit (Life Technologies).

    Techniques: Quantitative RT-PCR, Expressing, TUNEL Assay, Staining, Derivative Assay, ALP Assay

    Testosterone Modulation of AR Activity during MYC Activation (A–I) WT, K14MycER (Myc:), K14MycER AR-Shah , and K14MycER AR-TFM mice were treated as indicated. (A) Skin sections were stained with H E. (B and C) Skin sections were stained with the antibodies shown. (D) Quantitation of the SG differentiation compartment. (E) Quantitation of Ki67+ve sebocytes per SG. (F) qRT-PCR of Nucleolin (Nuc) mRNA levels relative to Gapdh . (G and H) Skin sections were stained with H E. Quantitation of SG differentiation compartment size is shown. (I) Schematic summary of the effect of testosterone on K14MycER mice treated with high-dose 4OHT. See Figure 1 N for stages in sebocytes differentiation. Increases in cell number are represented by +, ++, or +++, according to the strength of the effect. Reduction in cell number is represented by −. Three to six mice were examined per condition. Error bars represent SEM. ∗ p

    Journal: Cell Reports

    Article Title: c-MYC-Induced Sebaceous Gland Differentiation Is Controlled by an Androgen Receptor/p53 Axis

    doi: 10.1016/j.celrep.2013.01.013

    Figure Lengend Snippet: Testosterone Modulation of AR Activity during MYC Activation (A–I) WT, K14MycER (Myc:), K14MycER AR-Shah , and K14MycER AR-TFM mice were treated as indicated. (A) Skin sections were stained with H E. (B and C) Skin sections were stained with the antibodies shown. (D) Quantitation of the SG differentiation compartment. (E) Quantitation of Ki67+ve sebocytes per SG. (F) qRT-PCR of Nucleolin (Nuc) mRNA levels relative to Gapdh . (G and H) Skin sections were stained with H E. Quantitation of SG differentiation compartment size is shown. (I) Schematic summary of the effect of testosterone on K14MycER mice treated with high-dose 4OHT. See Figure 1 N for stages in sebocytes differentiation. Increases in cell number are represented by +, ++, or +++, according to the strength of the effect. Reduction in cell number is represented by −. Three to six mice were examined per condition. Error bars represent SEM. ∗ p

    Article Snippet: 2 μg of RNA was collected and treated with RQ1 DNase (Promega M610A) according to the manufacturer’s protocol. cDNA was prepared using Superscript III First-Strand Synthesis SuperMix for qRT-PCR Kit (Invitrogen 11752-250).

    Techniques: Activity Assay, Activation Assay, Mouse Assay, Staining, Quantitation Assay, Quantitative RT-PCR

    Effect of MYC and AR Activation on p53 Expression, DNA Double Strand Breaks, and Apoptosis (A–L) WT and K14MycER (Myc:) mice were treated as indicated. (A) Skin sections were labeled with antibodies to γ-H2AX with DAPI nuclear counterstain. (B, H, and K) Skin sections were labeled with antibodies to p53 (arrowheads indicate positive nuclei). (C) Skin sections were labeled with antibodies to cleaved caspase 3. (D–F) qRT-PCR of mRNA levels relative to Gapdh . (G, I, and L) Quantitation of p53+ve nuclei per unit area of skin (HF, SG, and overlying IFE). (J) qRT-PCR of p53 mRNA levels relative to Gapdh . Three to nine mice were examined per condition. Error bars represent SEM. ∗ p

    Journal: Cell Reports

    Article Title: c-MYC-Induced Sebaceous Gland Differentiation Is Controlled by an Androgen Receptor/p53 Axis

    doi: 10.1016/j.celrep.2013.01.013

    Figure Lengend Snippet: Effect of MYC and AR Activation on p53 Expression, DNA Double Strand Breaks, and Apoptosis (A–L) WT and K14MycER (Myc:) mice were treated as indicated. (A) Skin sections were labeled with antibodies to γ-H2AX with DAPI nuclear counterstain. (B, H, and K) Skin sections were labeled with antibodies to p53 (arrowheads indicate positive nuclei). (C) Skin sections were labeled with antibodies to cleaved caspase 3. (D–F) qRT-PCR of mRNA levels relative to Gapdh . (G, I, and L) Quantitation of p53+ve nuclei per unit area of skin (HF, SG, and overlying IFE). (J) qRT-PCR of p53 mRNA levels relative to Gapdh . Three to nine mice were examined per condition. Error bars represent SEM. ∗ p

    Article Snippet: 2 μg of RNA was collected and treated with RQ1 DNase (Promega M610A) according to the manufacturer’s protocol. cDNA was prepared using Superscript III First-Strand Synthesis SuperMix for qRT-PCR Kit (Invitrogen 11752-250).

    Techniques: Activation Assay, Expressing, Mouse Assay, Labeling, Quantitative RT-PCR, Quantitation Assay

    Effects of MYC Activation on SG Proliferation and Differentiation (A–Q) WT and K14MycER (Myc:) mouse telogen back skin was untreated, acetone-treated (vehicle), or 4OHT-treated, as indicated. (A, D, F, H, and L) Skin sections were stained with H E. (B, E, G, I, and M) Skin sections were stained with the antibodies shown. White bracket indicates expansion of AR-expressing sebocytes in (I). (C) Quantitation of SG differentiation compartment (average cross-sectional area occupied by differentiated sebocytes per SG) for the experiment described above. Red line represents average untreated WT measurement, and dashed red lines represent SEM. (J) Quantification of average size of individual differentiated sebocytes (cross-sectional area). (K) Average number of differentiated sebocytes per SG cross-section. (N) Quantitation of Ki67+ve sebocytes per SG. Red line represents untreated WT measurement. (O and P) qRT-PCR of Ki67 and Nucleolin mRNA levels relative to Gapdh . (Q) Schematic summary of changes relative to WT SG of treating K14MycER mice with low or high dose 4OHT. See Figure 1 N for stages in sebocytes differentiation. Increases in cell number are represented by +, ++, or +++, according to the strength of the effect. Reduction in cell number is represented by −. Three to nine mice were examined per condition. Error bars represent SEM #p

    Journal: Cell Reports

    Article Title: c-MYC-Induced Sebaceous Gland Differentiation Is Controlled by an Androgen Receptor/p53 Axis

    doi: 10.1016/j.celrep.2013.01.013

    Figure Lengend Snippet: Effects of MYC Activation on SG Proliferation and Differentiation (A–Q) WT and K14MycER (Myc:) mouse telogen back skin was untreated, acetone-treated (vehicle), or 4OHT-treated, as indicated. (A, D, F, H, and L) Skin sections were stained with H E. (B, E, G, I, and M) Skin sections were stained with the antibodies shown. White bracket indicates expansion of AR-expressing sebocytes in (I). (C) Quantitation of SG differentiation compartment (average cross-sectional area occupied by differentiated sebocytes per SG) for the experiment described above. Red line represents average untreated WT measurement, and dashed red lines represent SEM. (J) Quantification of average size of individual differentiated sebocytes (cross-sectional area). (K) Average number of differentiated sebocytes per SG cross-section. (N) Quantitation of Ki67+ve sebocytes per SG. Red line represents untreated WT measurement. (O and P) qRT-PCR of Ki67 and Nucleolin mRNA levels relative to Gapdh . (Q) Schematic summary of changes relative to WT SG of treating K14MycER mice with low or high dose 4OHT. See Figure 1 N for stages in sebocytes differentiation. Increases in cell number are represented by +, ++, or +++, according to the strength of the effect. Reduction in cell number is represented by −. Three to nine mice were examined per condition. Error bars represent SEM #p

    Article Snippet: 2 μg of RNA was collected and treated with RQ1 DNase (Promega M610A) according to the manufacturer’s protocol. cDNA was prepared using Superscript III First-Strand Synthesis SuperMix for qRT-PCR Kit (Invitrogen 11752-250).

    Techniques: Activation Assay, Staining, Expressing, Quantitation Assay, Quantitative RT-PCR, Mouse Assay

    Further Characterization of K14MycER Mice and Effects of MYC activation on AR Signaling, Related to Figure 2 (A) Scheme of experimental treatments. (B) Examples of differentiated SG compartment, (cross-sectional area occupied by differentiated sebocytes within yellow-dashed regions). (C) Ki67 labeling of K14MycER and WT control mice 4 days following treatment, quantified in Figure 2 N. (D–G) qRT-PCR of mRNA levels of Ar (D, E), Pcna (F), and K7 (G) relative to Gapdh . Error bars represent SEM. (H) AR-reporter luciferase assays in SebE6E7 human immortalized sebocytes. Data are means of 5 independent experiments ± SEM. NS = not significant, ∗ p

    Journal: Cell Reports

    Article Title: c-MYC-Induced Sebaceous Gland Differentiation Is Controlled by an Androgen Receptor/p53 Axis

    doi: 10.1016/j.celrep.2013.01.013

    Figure Lengend Snippet: Further Characterization of K14MycER Mice and Effects of MYC activation on AR Signaling, Related to Figure 2 (A) Scheme of experimental treatments. (B) Examples of differentiated SG compartment, (cross-sectional area occupied by differentiated sebocytes within yellow-dashed regions). (C) Ki67 labeling of K14MycER and WT control mice 4 days following treatment, quantified in Figure 2 N. (D–G) qRT-PCR of mRNA levels of Ar (D, E), Pcna (F), and K7 (G) relative to Gapdh . Error bars represent SEM. (H) AR-reporter luciferase assays in SebE6E7 human immortalized sebocytes. Data are means of 5 independent experiments ± SEM. NS = not significant, ∗ p

    Article Snippet: 2 μg of RNA was collected and treated with RQ1 DNase (Promega M610A) according to the manufacturer’s protocol. cDNA was prepared using Superscript III First-Strand Synthesis SuperMix for qRT-PCR Kit (Invitrogen 11752-250).

    Techniques: Mouse Assay, Activation Assay, Labeling, Quantitative RT-PCR, Luciferase

    Effect of Loss of AR Function during MYC Activation (A–H) WT and K14MycER mice (Myc:) were crossed with AR-Shah or AR-TFM mice and treated as indicated. (A and B) Skin sections were stained with H E. (C) Quantitation of size of SG differentiation compartment. (D and E) Immunostaining (brown) for antibodies indicated. (F) Quantitation of Ki67+ve sebocytes. (G) qRT-PCR of Nucleolin mRNA levels relative to Gapdh . (H) Schematic summary of the effects of AR loss of function. See Figure 1 N for stages in sebocyte differentiation. Increases in cell number are represented by +, ++, or +++, according to the strength of the effect. Reduction in cell number is represented by −. Three to seven mice were examined per condition. Error bars represent SEM. ∗ p

    Journal: Cell Reports

    Article Title: c-MYC-Induced Sebaceous Gland Differentiation Is Controlled by an Androgen Receptor/p53 Axis

    doi: 10.1016/j.celrep.2013.01.013

    Figure Lengend Snippet: Effect of Loss of AR Function during MYC Activation (A–H) WT and K14MycER mice (Myc:) were crossed with AR-Shah or AR-TFM mice and treated as indicated. (A and B) Skin sections were stained with H E. (C) Quantitation of size of SG differentiation compartment. (D and E) Immunostaining (brown) for antibodies indicated. (F) Quantitation of Ki67+ve sebocytes. (G) qRT-PCR of Nucleolin mRNA levels relative to Gapdh . (H) Schematic summary of the effects of AR loss of function. See Figure 1 N for stages in sebocyte differentiation. Increases in cell number are represented by +, ++, or +++, according to the strength of the effect. Reduction in cell number is represented by −. Three to seven mice were examined per condition. Error bars represent SEM. ∗ p

    Article Snippet: 2 μg of RNA was collected and treated with RQ1 DNase (Promega M610A) according to the manufacturer’s protocol. cDNA was prepared using Superscript III First-Strand Synthesis SuperMix for qRT-PCR Kit (Invitrogen 11752-250).

    Techniques: Activation Assay, Mouse Assay, Staining, Quantitation Assay, Immunostaining, Quantitative RT-PCR

    Loss of p53 Function during MYC Activation (A–K) Mice were treated as indicated. (A and J) Skin sections were stained with H E. (B) Quantitation of SG differentiation compartment. (C and J) Skin sections were stained with the antibodies shown. Arrowhead in (J) indicates sebocyte differentiation within the IFE. (D–I) qRT-PCR of mRNA levels relative to Gapdh . (K) Schematic summary of effects of loss of p53 and combined loss of p53 and the AR. See Figure 1 N for stages in sebocytes differentiation. Increases in cell number are represented by +, ++, or +++, according to the strength of the effect. Reduction in cell number is represented by −. Scale bars, 40 μm. Error bars represent SEM. Three to five mice were examined per condition, except n = 1 for triple cross. NS, not significant. ∗ p

    Journal: Cell Reports

    Article Title: c-MYC-Induced Sebaceous Gland Differentiation Is Controlled by an Androgen Receptor/p53 Axis

    doi: 10.1016/j.celrep.2013.01.013

    Figure Lengend Snippet: Loss of p53 Function during MYC Activation (A–K) Mice were treated as indicated. (A and J) Skin sections were stained with H E. (B) Quantitation of SG differentiation compartment. (C and J) Skin sections were stained with the antibodies shown. Arrowhead in (J) indicates sebocyte differentiation within the IFE. (D–I) qRT-PCR of mRNA levels relative to Gapdh . (K) Schematic summary of effects of loss of p53 and combined loss of p53 and the AR. See Figure 1 N for stages in sebocytes differentiation. Increases in cell number are represented by +, ++, or +++, according to the strength of the effect. Reduction in cell number is represented by −. Scale bars, 40 μm. Error bars represent SEM. Three to five mice were examined per condition, except n = 1 for triple cross. NS, not significant. ∗ p

    Article Snippet: 2 μg of RNA was collected and treated with RQ1 DNase (Promega M610A) according to the manufacturer’s protocol. cDNA was prepared using Superscript III First-Strand Synthesis SuperMix for qRT-PCR Kit (Invitrogen 11752-250).

    Techniques: Activation Assay, Mouse Assay, Staining, Quantitation Assay, Quantitative RT-PCR