tardbp gene expression (Qiagen)

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RNeasy FFPE Kit

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For purification of total RNA from formalin fixed paraffin embedded tissue sections Kit contents Qiagen RNeasy FFPE Kit 50 preps 14 to 30L Elution Volume FFPE Tissue Sample Total RNA Purification Spin Column Format Silica Technology Ideal for PCR qPCR Real time RT PCR Microarray Includes 50 RNeasy MinElute Spin Columns Collection Tubes Proteinase K RNase free DNase I DNase Booster Buffer RNase free Buffers RNase free Water Benefits Novel method to overcome formalin crosslinking Efficient release of RNA without compromising integrity Streamlined protocol providing RNA in just 85 minu

Catalog Number:

73504

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445

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RNeasy FFPE Kit

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Qiagen tardbp gene expression

tardbp gene expression - by Bioz Stars, 2020-05

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Images

1) Product Images from "Phosphorylated TDP-43 (pTDP-43) aggregates in the axial skeletal muscle of patients with sporadic and familial amyotrophic lateral sclerosis"

Article Title: Phosphorylated TDP-43 (pTDP-43) aggregates in the axial skeletal muscle of patients with sporadic and familial amyotrophic lateral sclerosis

Journal: Acta Neuropathologica Communications

doi: 10.1186/s40478-018-0528-y

). Three non-ALS, non-IBM samples with mild neurogenic atrophy in the muscle biopsy were also studied (“MSC1–3”). SQSTM1 and TARDBP were analyzed relative to the expression of two housekeeping genes ( 18 s , GAPDH ) and data shown are combined from two 96-well plates (4 replicates per sample and primer). There was a significant increase in relative normalized gene expression for both ALS and IBM samples ( a ), relative to controls (right) and this was most significant for SQSTM1 expression in ALS samples ( > 4-fold expression relative to controls, P

Figure Legend Snippet: ). Three non-ALS, non-IBM samples with mild neurogenic atrophy in the muscle biopsy were also studied (“MSC1–3”). SQSTM1 and TARDBP were analyzed relative to the expression of two housekeeping genes ( 18 s , GAPDH ) and data shown are combined from two 96-well plates (4 replicates per sample and primer). There was a significant increase in relative normalized gene expression for both ALS and IBM samples ( a ), relative to controls (right) and this was most significant for SQSTM1 expression in ALS samples ( > 4-fold expression relative to controls, P

Techniques Used: Expressing

2) Product Images from "Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies"

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

Journal: BMC Medical Research Methodology

doi: 10.1186/s12874-018-0628-1

Bland-Altman plots for investigating the level of agreement between RNA extraction kits. Each plot shows the differences between the two kits against the averages of the two kits. The lines represent the mean differences and upper and lower limits of agreement (LOA, mean differences ±1.96SD). a Comparison of RNA yield (ng/μl) of samples extracted with High Pure FFPE RNA Micro Kit and RNeasy® FFPE kit. b Comparison of purity (A260/A280) of samples extracted with High Pure FFPE RNA Micro kit and RNeasy® FFPE kit. c Comparison of RIN-values of samples extracted with High Pure FFPE RNA Micro kit and RNeasy® FFPE kit. d Comparison of RNA yield (ng/μl) of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit. e Comparison of purity (A260/A280) of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit. f Comparison of RIN-values of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit

Figure Legend Snippet: Bland-Altman plots for investigating the level of agreement between RNA extraction kits. Each plot shows the differences between the two kits against the averages of the two kits. The lines represent the mean differences and upper and lower limits of agreement (LOA, mean differences ±1.96SD). a Comparison of RNA yield (ng/μl) of samples extracted with High Pure FFPE RNA Micro Kit and RNeasy® FFPE kit. b Comparison of purity (A260/A280) of samples extracted with High Pure FFPE RNA Micro kit and RNeasy® FFPE kit. c Comparison of RIN-values of samples extracted with High Pure FFPE RNA Micro kit and RNeasy® FFPE kit. d Comparison of RNA yield (ng/μl) of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit. e Comparison of purity (A260/A280) of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit. f Comparison of RIN-values of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit

Techniques Used: RNA Extraction, Formalin-fixed Paraffin-Embedded

3) Product Images from "Nucleic acid extraction from formalin-fixed paraffin-embedded cancer cell line samples: a trade off between quantity and quality?"

Article Title: Nucleic acid extraction from formalin-fixed paraffin-embedded cancer cell line samples: a trade off between quantity and quality?

Journal: BMC Clinical Pathology

doi: 10.1186/s12907-016-0039-3

Investigating the relationship between RNA concentration and the input amount of tissue: Geometric mean increase in RNA concentration ( n = 6) when combining 5 μm sections in a linear fashion and associated standard error of mean, line of Y = X/5 represents a linear relationship a Qiagen AllPrep DNA/RNA FFPE kit, b Qiagen RNeasy, c Arcturus Paradise plus FFPE RNA isolation kit, d Maxwell 16 LEV RNA FFPE Purification kit

Figure Legend Snippet: Investigating the relationship between RNA concentration and the input amount of tissue: Geometric mean increase in RNA concentration ( n = 6) when combining 5 μm sections in a linear fashion and associated standard error of mean, line of Y = X/5 represents a linear relationship a Qiagen AllPrep DNA/RNA FFPE kit, b Qiagen RNeasy, c Arcturus Paradise plus FFPE RNA isolation kit, d Maxwell 16 LEV RNA FFPE Purification kit

Techniques Used: Concentration Assay, Formalin-fixed Paraffin-Embedded, Isolation, Purification

4) Product Images from "Bromodomain Protein BRD4 Is a Transcriptional Repressor of Autophagy and Lysosomal Function"

Article Title: Bromodomain Protein BRD4 Is a Transcriptional Repressor of Autophagy and Lysosomal Function

Journal: Molecular Cell

doi: 10.1016/j.molcel.2017.04.027

Starvation Leads to BRD4 Dissociation from Autophagy Gene Promoters (A and B) KP-4 cells were starved for 4 hr followed by chromatin immunoprecipitation (ChIP) assay with BRD4 antibody (A) and RT-qPCR analysis (B). (C) KP-4 cells infected with Cas9/hMOF sgRNA were subjected to ChIP assay with BRD4 antibody. (D) KP-4 cells were starved for 4 hr followed by ChIP assay with H4K16Ac antibody. (E and F) KP-4 cells infected with Cas9/SIRT1 sgRNA were starved for 4 hr followed by ChIP assay with BRD4 antibody (E) and RT-qPCR analysis (F). Western blot shows efficient SIRT1 depletion in Cas9/SIRT1 sgRNA-infected cells. (G and H) KP-4 cells infected with Cas9/AMPKα1 and α2 sgRNAs were starved for 4 hr followed by immunoprecipitation with DBC1 antibody (G) and RT-qPCR analysis (H). All data are shown as mean ± SD. In (A)–(F) and (H), n = 3 independent experiments. ∗ p

Figure Legend Snippet: Starvation Leads to BRD4 Dissociation from Autophagy Gene Promoters (A and B) KP-4 cells were starved for 4 hr followed by chromatin immunoprecipitation (ChIP) assay with BRD4 antibody (A) and RT-qPCR analysis (B). (C) KP-4 cells infected with Cas9/hMOF sgRNA were subjected to ChIP assay with BRD4 antibody. (D) KP-4 cells were starved for 4 hr followed by ChIP assay with H4K16Ac antibody. (E and F) KP-4 cells infected with Cas9/SIRT1 sgRNA were starved for 4 hr followed by ChIP assay with BRD4 antibody (E) and RT-qPCR analysis (F). Western blot shows efficient SIRT1 depletion in Cas9/SIRT1 sgRNA-infected cells. (G and H) KP-4 cells infected with Cas9/AMPKα1 and α2 sgRNAs were starved for 4 hr followed by immunoprecipitation with DBC1 antibody (G) and RT-qPCR analysis (H). All data are shown as mean ± SD. In (A)–(F) and (H), n = 3 independent experiments. ∗ p

Techniques Used: Chromatin Immunoprecipitation, Quantitative RT-PCR, Infection, Western Blot, Immunoprecipitation

BRD4 Represses Autophagy Gene Expression through G9a (A) Cell extracts from KP-4 cells were subjected to immunoprecipitation with G9a (upper) and BRD4 (lower) antibodies. (B) KP-4 cells were starved for 4 hr followed by immunoprecipitation with BRD4 antibody. (C–F) KP-4 cells were treated with 500 nM JQ1 for 9 hr (C and E). KP-4 cells harboring inducible control or BRD4 shRNA were treated with 500 ng/mL doxycycline (DOX) for 4 days (D and F). ChIP assays were performed using G9a (C and D) and H3K9diMe (E and F) antibodies. (G and H) KP-4 cells infected with shRNA targeting G9a were transfected with BRD4 siRNA followed by RT-qPCR (G) and western blot (H). (I) KP-4 cells overexpressing BRD4 were infected with shRNA targeting G9a. All data are shown as mean ± SD. In (C)–(G), n = 3 independent experiments. ∗ p

Figure Legend Snippet: BRD4 Represses Autophagy Gene Expression through G9a (A) Cell extracts from KP-4 cells were subjected to immunoprecipitation with G9a (upper) and BRD4 (lower) antibodies. (B) KP-4 cells were starved for 4 hr followed by immunoprecipitation with BRD4 antibody. (C–F) KP-4 cells were treated with 500 nM JQ1 for 9 hr (C and E). KP-4 cells harboring inducible control or BRD4 shRNA were treated with 500 ng/mL doxycycline (DOX) for 4 days (D and F). ChIP assays were performed using G9a (C and D) and H3K9diMe (E and F) antibodies. (G and H) KP-4 cells infected with shRNA targeting G9a were transfected with BRD4 siRNA followed by RT-qPCR (G) and western blot (H). (I) KP-4 cells overexpressing BRD4 were infected with shRNA targeting G9a. All data are shown as mean ± SD. In (C)–(G), n = 3 independent experiments. ∗ p

Techniques Used: Expressing, Immunoprecipitation, shRNA, Chromatin Immunoprecipitation, Infection, Transfection, Quantitative RT-PCR, Western Blot

BRD4 Is a Negative Regulator of Autophagy Gene Expression (A and B) KP-4 cells transfected with control or BRD4 siRNA were subjected to RNA-seq and gene ontology analyses (A) and RT-qPCR analysis (B). (C) RT-qPCR analysis of KP-4 cells treated with DMSO, 500 nM JQ1, 500 nM I-BET151, or 500 nM OTX015 for 9 hr. (D) RT-qPCR analysis of KP-4 cells treated with 500 nM JQ1 for the indicated time. (E) RT-qPCR analysis of KP-4 cells overexpressing BRD4. All data are shown as mean ± SD. In (A)–(D), n = 3 independent experiments; in (E), data are representative of two independent experiments performed in triplicate. ∗ p

Figure Legend Snippet: BRD4 Is a Negative Regulator of Autophagy Gene Expression (A and B) KP-4 cells transfected with control or BRD4 siRNA were subjected to RNA-seq and gene ontology analyses (A) and RT-qPCR analysis (B). (C) RT-qPCR analysis of KP-4 cells treated with DMSO, 500 nM JQ1, 500 nM I-BET151, or 500 nM OTX015 for 9 hr. (D) RT-qPCR analysis of KP-4 cells treated with 500 nM JQ1 for the indicated time. (E) RT-qPCR analysis of KP-4 cells overexpressing BRD4. All data are shown as mean ± SD. In (A)–(D), n = 3 independent experiments; in (E), data are representative of two independent experiments performed in triplicate. ∗ p

Techniques Used: Expressing, Transfection, RNA Sequencing Assay, Quantitative RT-PCR

BRD4 Knockdown Sustains mTOR Activity during Starvation and Confers Resistance to Starvation-Induced Cell Death (A and B) KP-4 cells transfected with BRD4 siRNA were starved of amino acids (A). Cells pre-treated with CQ (10 μM, 4 hr) were subjected to amino acid starvation for 2 hr in the presence of CQ (B). (C) KP-4 cells infected with Cas9/ATG5 sgRNA were transfected with BRD4 siRNA and subjected to amino acid starvation for 2 hr. (D and E) KP-4 cells infected with Cas9/ATG5 sgRNA were transfected with BRD4 siRNA. Following 48 hr starvation, percentage of subG1 cells (D) and cell number (E) were determined (n = 3 independent experiments). (F and G) KP-4 cells transfected with BRD4 siRNA were starved for 48 hr in the presence or absence of 10 μM CQ. Percentage of dead (F) and surviving (G) cells was determined by trypan blue exclusion test (n = 4 independent experiments). All data are shown as mean ± SD. ∗ p

Figure Legend Snippet: BRD4 Knockdown Sustains mTOR Activity during Starvation and Confers Resistance to Starvation-Induced Cell Death (A and B) KP-4 cells transfected with BRD4 siRNA were starved of amino acids (A). Cells pre-treated with CQ (10 μM, 4 hr) were subjected to amino acid starvation for 2 hr in the presence of CQ (B). (C) KP-4 cells infected with Cas9/ATG5 sgRNA were transfected with BRD4 siRNA and subjected to amino acid starvation for 2 hr. (D and E) KP-4 cells infected with Cas9/ATG5 sgRNA were transfected with BRD4 siRNA. Following 48 hr starvation, percentage of subG1 cells (D) and cell number (E) were determined (n = 3 independent experiments). (F and G) KP-4 cells transfected with BRD4 siRNA were starved for 48 hr in the presence or absence of 10 μM CQ. Percentage of dead (F) and surviving (G) cells was determined by trypan blue exclusion test (n = 4 independent experiments). All data are shown as mean ± SD. ∗ p

Techniques Used: Activity Assay, Transfection, Infection

BRD4 Knockdown Enhances Lysosomal Function (A) RT-qPCR analysis of KP-4 cells transfected with control or BRD4 siRNA. (B–D) KP-4 cells transfected with BRD4 siRNA were subjected to western blot analysis with antibodies against lysosomal proteins (B) and stained with LAMP1 antibody (C), LysoTracker Red (100 nM, 2 hr) (D, upper panels), and Magic Red CTSB (1 hr) (D, lower panels). Area of LAMP1 + , LysoTracker + , and Magic Red CTSB + area normalized to cell number is shown (C, CON: n = 115 cells, BRD4: n = 130 cells; D upper, CON: n = 66 cells, BRD4 1: n = 52 cells, BRD4 2: n = 50 cells; D lower, CON: n = 164 cells, BRD4 1: n = 109 cells, BRD4 2: n = 53 cells). Scale bars, 50 μm. (E) Hexosaminidase activity was measured using lysates from control and BRD4 knockdown KP-4 cells. (F and G) RT-qPCR analysis of TY-82 cells transfected with NUT siRNA for 72 hr (F) or treated with 500 nM JQ1 for 9 hr (G). (H) KP-4 cells were transfected with BRD4 and/or MiT/TFE (TFEB, TFE3, MITF) siRNAs and treated with 10 μM CQ for 4 hr. All data are shown as mean ± SD. In (A) and (F), n = 3 independent experiments. In (E), n = 4 independent experiments. In (G), data are representative of two independent experiments performed in triplicate. ∗ p

Figure Legend Snippet: BRD4 Knockdown Enhances Lysosomal Function (A) RT-qPCR analysis of KP-4 cells transfected with control or BRD4 siRNA. (B–D) KP-4 cells transfected with BRD4 siRNA were subjected to western blot analysis with antibodies against lysosomal proteins (B) and stained with LAMP1 antibody (C), LysoTracker Red (100 nM, 2 hr) (D, upper panels), and Magic Red CTSB (1 hr) (D, lower panels). Area of LAMP1 + , LysoTracker + , and Magic Red CTSB + area normalized to cell number is shown (C, CON: n = 115 cells, BRD4: n = 130 cells; D upper, CON: n = 66 cells, BRD4 1: n = 52 cells, BRD4 2: n = 50 cells; D lower, CON: n = 164 cells, BRD4 1: n = 109 cells, BRD4 2: n = 53 cells). Scale bars, 50 μm. (E) Hexosaminidase activity was measured using lysates from control and BRD4 knockdown KP-4 cells. (F and G) RT-qPCR analysis of TY-82 cells transfected with NUT siRNA for 72 hr (F) or treated with 500 nM JQ1 for 9 hr (G). (H) KP-4 cells were transfected with BRD4 and/or MiT/TFE (TFEB, TFE3, MITF) siRNAs and treated with 10 μM CQ for 4 hr. All data are shown as mean ± SD. In (A) and (F), n = 3 independent experiments. In (E), n = 4 independent experiments. In (G), data are representative of two independent experiments performed in triplicate. ∗ p

Techniques Used: Quantitative RT-PCR, Transfection, Western Blot, Staining, Activity Assay

BRD4 Silencing Enhances Autophagic Flux (A) Drosophila S2R + cells expressing GFP-LC3 were transfected with double-stranded RNA (dsRNA) targeting control luciferase (Luc) or Fs(1)h. (B and C) KP-4 cells transfected with control or BRD4 siRNA for 72 hr were subjected to western blot analysis (B) and stained for LC3B (C). The number of LC3 puncta normalized to cell number is shown. CON: n = 94 cells, BRD4 1: n = 97 cells, BRD4 2: n = 74 cells. Scale bars, 50 μm. (D) Immunohistochemistry of small intestinal sections from transgenic mice harboring inducible renilla luciferase or BRD4 shRNA. Sections were stained for LC3 (upper) and BRD4 (lower). Cytoplasmic signal in BRD4 panels is due to non-specific staining. Scale bars, 50 μm. (E) KP-4 cells transfected with BRD4 siRNA were treated with 10 μM CQ for 4 hr. (F) KP-4 cells transfected with BRD4 siRNA were stained for WIPI2. The number of WIPI2 puncta normalized to cell number is shown. CON: n = 119 cells, BRD4 1: n = 107 cells, BRD4 2: n = 109 cells. Scale bars, 20 μm. (G) KP-4 cells stably expressing RFP-GFP-LC3 were transfected with BRD4 siRNA. Scale bars, 50 μm. (H) KP-4 cells were treated with 500 nM JQ1 for 9 hr in the presence or absence of CQ (10 μM, 4 hr). (I) KP-4 cells overexpressing BRD4 were treated with 10 μM CQ for 4 hr. (J) TY-82 cells transfected with NUT siRNA for 5 days were treated with 10 μM CQ for 8 hr. BRD4-NUT was detected using NUT antibody. All data are shown as mean ± SD. ∗ p

Figure Legend Snippet: BRD4 Silencing Enhances Autophagic Flux (A) Drosophila S2R + cells expressing GFP-LC3 were transfected with double-stranded RNA (dsRNA) targeting control luciferase (Luc) or Fs(1)h. (B and C) KP-4 cells transfected with control or BRD4 siRNA for 72 hr were subjected to western blot analysis (B) and stained for LC3B (C). The number of LC3 puncta normalized to cell number is shown. CON: n = 94 cells, BRD4 1: n = 97 cells, BRD4 2: n = 74 cells. Scale bars, 50 μm. (D) Immunohistochemistry of small intestinal sections from transgenic mice harboring inducible renilla luciferase or BRD4 shRNA. Sections were stained for LC3 (upper) and BRD4 (lower). Cytoplasmic signal in BRD4 panels is due to non-specific staining. Scale bars, 50 μm. (E) KP-4 cells transfected with BRD4 siRNA were treated with 10 μM CQ for 4 hr. (F) KP-4 cells transfected with BRD4 siRNA were stained for WIPI2. The number of WIPI2 puncta normalized to cell number is shown. CON: n = 119 cells, BRD4 1: n = 107 cells, BRD4 2: n = 109 cells. Scale bars, 20 μm. (G) KP-4 cells stably expressing RFP-GFP-LC3 were transfected with BRD4 siRNA. Scale bars, 50 μm. (H) KP-4 cells were treated with 500 nM JQ1 for 9 hr in the presence or absence of CQ (10 μM, 4 hr). (I) KP-4 cells overexpressing BRD4 were treated with 10 μM CQ for 4 hr. (J) TY-82 cells transfected with NUT siRNA for 5 days were treated with 10 μM CQ for 8 hr. BRD4-NUT was detected using NUT antibody. All data are shown as mean ± SD. ∗ p

Techniques Used: Expressing, Transfection, Luciferase, Western Blot, Staining, Immunohistochemistry, Transgenic Assay, Mouse Assay, shRNA, Stable Transfection

$Effect of BRD4 Silencing on Stimulus-Dependent and Selective Autophagy (A–F) Cells transfected with BRD4 siRNA were starved for 1–5 hr (KP-4 cells, A), treated with 500 nM rapamycin for 24 hr (KP-4 cells, B), starved of glucose for 4 hr (KP-4 cells, C), cultured under hypoxic (1% O 2 ) conditions for 48 hr (SUIT2 cells, D), treated with 100 mM Trehalose for 4 hr (KP-4 cells, E), or treated with 500 nM 4-Hydroxytamoxifen (4-OHT) for 48 hr (IMR90 ER-HRas G12V cells, F). (G) KP-4 cells harboring rtTA and Tre-tight-HTT Q94-CFP were transfected with BRD4 siRNA. At 12 hr after transfection, cells were treated with 1 μg/mL DOX for 10 hr. At 48 hr after removal of DOX, cells were separated into Triton X-100 soluble and insoluble fractions. (H) KP-4 cells transfected with BRD4 siRNA were infected with Salmonella enterica serovar Typhimurium. The number of Salmonella was determined by performing colony-forming unit assays at 2, 6, and 8 hr after infection and normalized to the numbers at 2 hr. Data are shown as mean ± SEM; n = 4 independent experiments. (I) KP-4 cells expressing YFP-parkin were transfected with BRD4 siRNA followed by treatment with 1 μM Antimycin A and 1 μM Oligomycin for 8 hr. See also Figure S6 .$
Figure Legend Snippet: Effect of BRD4 Silencing on Stimulus-Dependent and Selective Autophagy (A–F) Cells transfected with BRD4 siRNA were starved for 1–5 hr (KP-4 cells, A), treated with 500 nM rapamycin for 24 hr (KP-4 cells, B), starved of glucose for 4 hr (KP-4 cells, C), cultured under hypoxic (1% O 2 ) conditions for 48 hr (SUIT2 cells, D), treated with 100 mM Trehalose for 4 hr (KP-4 cells, E), or treated with 500 nM 4-Hydroxytamoxifen (4-OHT) for 48 hr (IMR90 ER-HRas G12V cells, F). (G) KP-4 cells harboring rtTA and Tre-tight-HTT Q94-CFP were transfected with BRD4 siRNA. At 12 hr after transfection, cells were treated with 1 μg/mL DOX for 10 hr. At 48 hr after removal of DOX, cells were separated into Triton X-100 soluble and insoluble fractions. (H) KP-4 cells transfected with BRD4 siRNA were infected with Salmonella enterica serovar Typhimurium. The number of Salmonella was determined by performing colony-forming unit assays at 2, 6, and 8 hr after infection and normalized to the numbers at 2 hr. Data are shown as mean ± SEM; n = 4 independent experiments. (I) KP-4 cells expressing YFP-parkin were transfected with BRD4 siRNA followed by treatment with 1 μM Antimycin A and 1 μM Oligomycin for 8 hr. See also Figure S6 .

Techniques Used: Transfection, Cell Culture, Infection, Expressing

5) Product Images from "Comparison of triple-negative breast cancer molecular subtyping using RNA from matched fresh-frozen versus formalin-fixed paraffin-embedded tissue"

Article Title: Comparison of triple-negative breast cancer molecular subtyping using RNA from matched fresh-frozen versus formalin-fixed paraffin-embedded tissue

Journal: BMC Cancer

doi: 10.1186/s12885-017-3237-1

TNBC molecular subtype concordance between matched FF and FFPE samples processed on microarray and RNA-seq improves with increased prediction confidence. a Scatterplot shows TNBC subtype accuracy between microarray and RNA-seq as a function of prediction confidence in the TCGA breast (BRCA) cohort. b Plot shows RNA-seq prediction accuracy by confidence score. Vertical line cutoff demarks the prediction confidence score generating 95% concordance between platforms. c Scatterplot shows the concordance between microarray and RNA-seq platforms by strength of correlation to a subtype (prediction score)

Figure Legend Snippet: TNBC molecular subtype concordance between matched FF and FFPE samples processed on microarray and RNA-seq improves with increased prediction confidence. a Scatterplot shows TNBC subtype accuracy between microarray and RNA-seq as a function of prediction confidence in the TCGA breast (BRCA) cohort. b Plot shows RNA-seq prediction accuracy by confidence score. Vertical line cutoff demarks the prediction confidence score generating 95% concordance between platforms. c Scatterplot shows the concordance between microarray and RNA-seq platforms by strength of correlation to a subtype (prediction score)

Techniques Used: Formalin-fixed Paraffin-Embedded, Microarray, RNA Sequencing Assay

MiSeq and HiSeq platform mapped read comparison from FF- and FFPE-derived RNA sequences. a Barplot depicts the percentage of mapped reads that are on-target, or off-target (intronic and intergenic) for FF and FFPE samples processed on MiSeq and HiSeq platforms. b Beeswarm box plot shows mapped reads (%) form individual FF ( blue ) and FFPE ( red ) samples processed on the HiSeq

Figure Legend Snippet: MiSeq and HiSeq platform mapped read comparison from FF- and FFPE-derived RNA sequences. a Barplot depicts the percentage of mapped reads that are on-target, or off-target (intronic and intergenic) for FF and FFPE samples processed on MiSeq and HiSeq platforms. b Beeswarm box plot shows mapped reads (%) form individual FF ( blue ) and FFPE ( red ) samples processed on the HiSeq

Techniques Used: Formalin-fixed Paraffin-Embedded, Derivative Assay

6) Product Images from "CDK1 and CCNB1 as potential diagnostic markers of rhabdomyosarcoma: validation following bioinformatics analysis"

Article Title: CDK1 and CCNB1 as potential diagnostic markers of rhabdomyosarcoma: validation following bioinformatics analysis

Journal: BMC Medical Genomics

doi: 10.1186/s12920-019-0645-x

Validation of 10 hub genes by RT-PCR. mRNA expression levels of 10 hub genes were detected in RMS patient tissue and normal striated muscle tissue. Each point represents an individual subject. * p

Figure Legend Snippet: Validation of 10 hub genes by RT-PCR. mRNA expression levels of 10 hub genes were detected in RMS patient tissue and normal striated muscle tissue. Each point represents an individual subject. * p

Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing

7) Product Images from "Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies"

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

Journal: BMC Medical Research Methodology

doi: 10.1186/s12874-018-0628-1

Techniques Used: RNA Extraction, Formalin-fixed Paraffin-Embedded

8) Product Images from "Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies"

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

Journal: BMC Medical Research Methodology

doi: 10.1186/s12874-018-0628-1

Techniques Used: RNA Extraction, Formalin-fixed Paraffin-Embedded

9) Product Images from "Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies"

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

Journal: BMC Medical Research Methodology

doi: 10.1186/s12874-018-0628-1

Techniques Used: RNA Extraction, Formalin-fixed Paraffin-Embedded

10) Product Images from "Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies"

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

Journal: BMC Medical Research Methodology

doi: 10.1186/s12874-018-0628-1

Techniques Used: RNA Extraction, Formalin-fixed Paraffin-Embedded

11) Product Images from "Upregulation of P2Y2R, Active uPA, and PAI-1 Are Essential Components of Hantavirus Cardiopulmonary Syndrome"

Article Title: Upregulation of P2Y2R, Active uPA, and PAI-1 Are Essential Components of Hantavirus Cardiopulmonary Syndrome

Journal: Frontiers in Cellular and Infection Microbiology

doi: 10.3389/fcimb.2018.00169

P2Y 2 R expression is significantly upregulated in HCPS compared to controls. RNA was extracted from lung tissues samples with the Qiagen RNeasy FFPE kit (cat# 73504) and quantified with a Thermo Fisher Scientific Nanodrop. P2Y 2 R expression levels were measured by the TaqMan assay using a WT P2Y 2 R plasmid as standard. P2Y 2 R copy #s were normalized to total RNA extracted from the embedded tissues. Each sample slice was measured in triplicate. (A) The plot of P2Y 2 R for each HCPS patient, shown on the x-axis as H1-H22. Each data point per patient represents a triplicate measurement of P2Y 2 R expression in a 10 μm slice cut from the FFPE blocks. The number of segments analyzed is shown for each case above the replicate data points (e.g., n = 11 for H1). Ninety-seven distinct segments from all cases were analyzed. Lung tissues from two decedents (H4, and H9) were not available. The error bars represent the median and range. For all HCPS samples, the minimum, median, maximum and mean values were 0.0055, 2.10, 24.88, and 4.1 ± 0.48 (SEM). (B,C) Plot of P2Y 2 R for pneumonia (P) and gunshot wound cases (GSW). The experimental conditions are similar to those described for HCPS. Cases for which no lung tissue samples were available were excluded from the graph. For all pneumonia tissue segment samples ( n = 39), the minimum, median, maximum and mean values were 0.0005, 0.07, 4.4, 0.54 ± 0.16. For all GSW samples ( n = 21), the minimum, median, maximum and mean values were 0.0052, 0.18, 2.4, and 0.46 ± 0.10 (SEM).

Figure Legend Snippet: P2Y 2 R expression is significantly upregulated in HCPS compared to controls. RNA was extracted from lung tissues samples with the Qiagen RNeasy FFPE kit (cat# 73504) and quantified with a Thermo Fisher Scientific Nanodrop. P2Y 2 R expression levels were measured by the TaqMan assay using a WT P2Y 2 R plasmid as standard. P2Y 2 R copy #s were normalized to total RNA extracted from the embedded tissues. Each sample slice was measured in triplicate. (A) The plot of P2Y 2 R for each HCPS patient, shown on the x-axis as H1-H22. Each data point per patient represents a triplicate measurement of P2Y 2 R expression in a 10 μm slice cut from the FFPE blocks. The number of segments analyzed is shown for each case above the replicate data points (e.g., n = 11 for H1). Ninety-seven distinct segments from all cases were analyzed. Lung tissues from two decedents (H4, and H9) were not available. The error bars represent the median and range. For all HCPS samples, the minimum, median, maximum and mean values were 0.0055, 2.10, 24.88, and 4.1 ± 0.48 (SEM). (B,C) Plot of P2Y 2 R for pneumonia (P) and gunshot wound cases (GSW). The experimental conditions are similar to those described for HCPS. Cases for which no lung tissue samples were available were excluded from the graph. For all pneumonia tissue segment samples ( n = 39), the minimum, median, maximum and mean values were 0.0005, 0.07, 4.4, 0.54 ± 0.16. For all GSW samples ( n = 21), the minimum, median, maximum and mean values were 0.0052, 0.18, 2.4, and 0.46 ± 0.10 (SEM).

Techniques Used: Expressing, Formalin-fixed Paraffin-Embedded, TaqMan Assay, Plasmid Preparation

12) Product Images from "Embryonic Stem Cell-Like Subpopulations in Venous Malformation"

Article Title: Embryonic Stem Cell-Like Subpopulations in Venous Malformation

Journal: Frontiers in Medicine

doi: 10.3389/fmed.2017.00162

3,3′-Diaminobenzidine immunohistochemical-stained images demonstrating the expression of Nanog [ (A,B) , red], pSTAT3 [ (C,D) , brown], OCT4 [ (E,F) , brown], SOX2 [ (G,H) , brown], SALL4 [ (I,J) , brown], and CD44 [ (K,L) , brown] in subcutaneous venous malformation (SCVM) (A,C,E,G,I,K) and intramuscular venous malformation (IMVM) (B,D,F,H,J,L) . Endothelial staining of all six embryonic stem cell markers was present on the endothelium within both SCVM and IMVM samples. Nanog (A,B) , pSTAT3 (C,D) , SOX2 (G,H) , and CD44 (K,L) were also expressed on cells ( arrowheads ) away from the endothelium in both SCVM and IMVM samples. Nuclei were counterstained with hematoxylin (blue). Original magnification: 400×.

Figure Legend Snippet: 3,3′-Diaminobenzidine immunohistochemical-stained images demonstrating the expression of Nanog [ (A,B) , red], pSTAT3 [ (C,D) , brown], OCT4 [ (E,F) , brown], SOX2 [ (G,H) , brown], SALL4 [ (I,J) , brown], and CD44 [ (K,L) , brown] in subcutaneous venous malformation (SCVM) (A,C,E,G,I,K) and intramuscular venous malformation (IMVM) (B,D,F,H,J,L) . Endothelial staining of all six embryonic stem cell markers was present on the endothelium within both SCVM and IMVM samples. Nanog (A,B) , pSTAT3 (C,D) , SOX2 (G,H) , and CD44 (K,L) were also expressed on cells ( arrowheads ) away from the endothelium in both SCVM and IMVM samples. Nuclei were counterstained with hematoxylin (blue). Original magnification: 400×.

Techniques Used: Immunohistochemistry, Staining, Expressing

Representative hematoxylin and eosin stained subcutaneous venous malformation (SCVM) (A) and intramuscular venous malformation (IMVM) (B) sections demonstrating the characteristic ectatic venous channels. Representative sections of SCVM (C) and IMVM (D) showing minimal staining for D2-40 [ (C,D) , brown]. Nuclei were counterstained with hematoxylin [ (A–D) , blue]. Original magnifications: 400× (A,B) and 100× (C,D) .

Figure Legend Snippet: Representative hematoxylin and eosin stained subcutaneous venous malformation (SCVM) (A) and intramuscular venous malformation (IMVM) (B) sections demonstrating the characteristic ectatic venous channels. Representative sections of SCVM (C) and IMVM (D) showing minimal staining for D2-40 [ (C,D) , brown]. Nuclei were counterstained with hematoxylin [ (A–D) , blue]. Original magnifications: 400× (A,B) and 100× (C,D) .

Techniques Used: Staining

Representative immunofluorescent immunohistochemical-stained sections of subcutaneous venous malformation (SCVM) (A) and intramuscular venous malformation (IMVM) (B) samples, demonstrating the endothelium consisted of CD34 + (green)/ERG − (red) ( long arrows ), ERG + (red)/CD34 − (green) endothelium ( arrowheads ), and CD34 + (red)/ERG + (red) ( short arrows ) phenotypes. The CD34 + (green) endothelium expressed Nanog (red, arrows ) in SCVM (C) and IMVM (D) lesions with cells away from the endothelium also expressing Nanog (red, arrowheads ) within SCVM (C) and IMVM (D) lesions. The CD34 + (green) endothelium expressed pSTAT3 (red, arrows ) in both SCVM (E) and IMVM (F) lesions. Cells away from the endothelium also expressed pSTAT3 (red, arrowheads ) within SCVM (E) and IMVM (F) lesions. The ERG + (red) endothelium also expressed OCT4 (green, arrows ) in both SCVM (G) and IMVM (H) lesions. The CD34 + (green) endothelium expressed SOX2 (red, arrows ) in SCVM (I) and IMVM (J) lesions. Cells away from the endothelium also expressed SOX2 (red, arrowheads ) in SCVM (I) and IMVM (J) lesions. The ERG + endothelium (red) expressed SALL4 (green, arrows ) in SCVM (K) and IMVM (L) lesions. Dual staining of with SOX2 and SALL4 demonstrated the SALL4 + [ (M,N) , green] endothelial population expressed SOX2 [ (M,N) , red] in both SCVM (M) and IMVM (N) . The ERG + endothelium (red) expressed CD44 (green, arrows ) in SCVM (O) and IMVM (P) lesions with cells away from the endothelium also expressing CD44 (green, arrowheads ) in SCVM (O) and IMVM (P) . Cells outside of the endothelium in both SCVM (Q) and IMVM (R) co-expressed Nanog [ (Q,R) , red] and CD44 [ (Q,R) , green]. Cell nuclei were counterstained with 4′,6′-diamidino-2-phenylindole [ (A–R) , blue]. Scale bars: 20 µm.

Figure Legend Snippet: Representative immunofluorescent immunohistochemical-stained sections of subcutaneous venous malformation (SCVM) (A) and intramuscular venous malformation (IMVM) (B) samples, demonstrating the endothelium consisted of CD34 + (green)/ERG − (red) ( long arrows ), ERG + (red)/CD34 − (green) endothelium ( arrowheads ), and CD34 + (red)/ERG + (red) ( short arrows ) phenotypes. The CD34 + (green) endothelium expressed Nanog (red, arrows ) in SCVM (C) and IMVM (D) lesions with cells away from the endothelium also expressing Nanog (red, arrowheads ) within SCVM (C) and IMVM (D) lesions. The CD34 + (green) endothelium expressed pSTAT3 (red, arrows ) in both SCVM (E) and IMVM (F) lesions. Cells away from the endothelium also expressed pSTAT3 (red, arrowheads ) within SCVM (E) and IMVM (F) lesions. The ERG + (red) endothelium also expressed OCT4 (green, arrows ) in both SCVM (G) and IMVM (H) lesions. The CD34 + (green) endothelium expressed SOX2 (red, arrows ) in SCVM (I) and IMVM (J) lesions. Cells away from the endothelium also expressed SOX2 (red, arrowheads ) in SCVM (I) and IMVM (J) lesions. The ERG + endothelium (red) expressed SALL4 (green, arrows ) in SCVM (K) and IMVM (L) lesions. Dual staining of with SOX2 and SALL4 demonstrated the SALL4 + [ (M,N) , green] endothelial population expressed SOX2 [ (M,N) , red] in both SCVM (M) and IMVM (N) . The ERG + endothelium (red) expressed CD44 (green, arrows ) in SCVM (O) and IMVM (P) lesions with cells away from the endothelium also expressing CD44 (green, arrowheads ) in SCVM (O) and IMVM (P) . Cells outside of the endothelium in both SCVM (Q) and IMVM (R) co-expressed Nanog [ (Q,R) , red] and CD44 [ (Q,R) , green]. Cell nuclei were counterstained with 4′,6′-diamidino-2-phenylindole [ (A–R) , blue]. Scale bars: 20 µm.

Techniques Used: Immunohistochemistry, Staining, Expressing

Log 10 relative expression of OCT4, STAT3, and CD44 (A) and SOX2, Nanog, and SALL4 (B) mRNA transcripts in three subcutaneous venous malformation (SCVM) and three intramuscular venous malformation (IMVM) samples analyzed by NanoString (A) and RT-qPCR (B) analyses. Expression is depicted relative to the housekeeping gene GAPDH. OCT4 was detected in two SCVM and two IMVM samples (A) . STAT3 and CD44 (A) and SOX2 and SALL4 (B) were detected in all three samples. Nanog was detected in all three SCVM samples and two out of three IMVM samples (B) .

Figure Legend Snippet: Log 10 relative expression of OCT4, STAT3, and CD44 (A) and SOX2, Nanog, and SALL4 (B) mRNA transcripts in three subcutaneous venous malformation (SCVM) and three intramuscular venous malformation (IMVM) samples analyzed by NanoString (A) and RT-qPCR (B) analyses. Expression is depicted relative to the housekeeping gene GAPDH. OCT4 was detected in two SCVM and two IMVM samples (A) . STAT3 and CD44 (A) and SOX2 and SALL4 (B) were detected in all three samples. Nanog was detected in all three SCVM samples and two out of three IMVM samples (B) .

Techniques Used: Expressing, Quantitative RT-PCR

13) Product Images from "Felis catus papillomavirus type-2 E6 binds to E6AP, promotes E6AP/p53 binding and enhances p53 proteasomal degradation"

Article Title: Felis catus papillomavirus type-2 E6 binds to E6AP, promotes E6AP/p53 binding and enhances p53 proteasomal degradation

Journal: Scientific Reports

doi: 10.1038/s41598-018-35723-7

FcaPV-2 E6 mRNA levels, expression and localization of p53 and E6AP in feline oral SCC cell lines. ( a ) SCCF2 and SCCF3 cells were subjected to RNA extraction, reverse transcription and Real time quantitative PCR (qPCR) to analyse FcaPV-2 E6 gene expression. Results were normalised for β2-microglobulin expression. The graph shows the results as mean +/− standard deviations (SD) of five independent experiments where the cell line with higher E6 expression levels (SCCF3) (P = 0.0062) was arbitrarily set as calibrator. ( b ) Cells were analysed by Real-time qPCR for p53 gene expression and results normalized as above. The graph represents mean +/− SD from three independent experiment and demonstrates p53 down-regulation in SCCF3 compared to SCCF2 set as calibrator (P = 0.00). Representative WB gels for p53 and E6AP are also shown. Stripping and reprobing with β-actin confirmed equal protein loadings. ( c ) SCCF2 and SCCF3 cells were grown on coverslips and subjected to double IF staining for p53 (green fluorescence) and E6AP (red fluorescence), nuclei were counterstained with DAPI. Representative fields showing nuclear or cytoplasmic/perinuclear p53 expression in SCCF2 and undetectable p53 staining in SCCF3 are illustrated. White arrows indicate perinuclear co-localization (yellow fluorescence) of the two proteins in a sub-set of SCCF2 cells in the merge panel.

Figure Legend Snippet: FcaPV-2 E6 mRNA levels, expression and localization of p53 and E6AP in feline oral SCC cell lines. ( a ) SCCF2 and SCCF3 cells were subjected to RNA extraction, reverse transcription and Real time quantitative PCR (qPCR) to analyse FcaPV-2 E6 gene expression. Results were normalised for β2-microglobulin expression. The graph shows the results as mean +/− standard deviations (SD) of five independent experiments where the cell line with higher E6 expression levels (SCCF3) (P = 0.0062) was arbitrarily set as calibrator. ( b ) Cells were analysed by Real-time qPCR for p53 gene expression and results normalized as above. The graph represents mean +/− SD from three independent experiment and demonstrates p53 down-regulation in SCCF3 compared to SCCF2 set as calibrator (P = 0.00). Representative WB gels for p53 and E6AP are also shown. Stripping and reprobing with β-actin confirmed equal protein loadings. ( c ) SCCF2 and SCCF3 cells were grown on coverslips and subjected to double IF staining for p53 (green fluorescence) and E6AP (red fluorescence), nuclei were counterstained with DAPI. Representative fields showing nuclear or cytoplasmic/perinuclear p53 expression in SCCF2 and undetectable p53 staining in SCCF3 are illustrated. White arrows indicate perinuclear co-localization (yellow fluorescence) of the two proteins in a sub-set of SCCF2 cells in the merge panel.

Techniques Used: Expressing, RNA Extraction, Real-time Polymerase Chain Reaction, Western Blot, Stripping Membranes, Staining, Fluorescence

14) Product Images from "Evaluation and Validation of Total RNA Extraction Methods for MicroRNA Expression Analyses in Formalin-Fixed, Paraffin-Embedded Tissues"

Article Title: Evaluation and Validation of Total RNA Extraction Methods for MicroRNA Expression Analyses in Formalin-Fixed, Paraffin-Embedded Tissues

Journal:

doi: 10.2353/jmoldx.2008.070153

qRT-PCR analysis of RNA samples isolated with the RecoverAll or RNeasy kit. Total RNA was extracted in triplicate from five different FFPE tissue blocks using the indicated method. Each RNA sample was analyzed in duplicate using qRT-PCR assays specific

Figure Legend Snippet: qRT-PCR analysis of RNA samples isolated with the RecoverAll or RNeasy kit. Total RNA was extracted in triplicate from five different FFPE tissue blocks using the indicated method. Each RNA sample was analyzed in duplicate using qRT-PCR assays specific

Techniques Used: Quantitative RT-PCR, Isolation, Formalin-fixed Paraffin-Embedded

15) Product Images from "Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies"

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

Journal: BMC Medical Research Methodology

doi: 10.1186/s12874-018-0628-1

Techniques Used: RNA Extraction, Formalin-fixed Paraffin-Embedded

16) Product Images from "Transcriptomic Profiling of Human Peritumoral Neocortex Tissues Revealed Genes Possibly Involved in Tumor-Induced Epilepsy"

Article Title: Transcriptomic Profiling of Human Peritumoral Neocortex Tissues Revealed Genes Possibly Involved in Tumor-Induced Epilepsy

Journal: PLoS ONE

doi: 10.1371/journal.pone.0056077

Bioanalyzer profiles of FFPE human peritumoral tissue total RNA. The total RNA was extracted from FFPE peritumoral tissues using Qiagen RNA extraction kit for FFPE samples or using Qiagen RNeasy Mini kit for human peripheral blood mononuclear cells (PBMCs). The upper panel shows the Bioanalyzer results of two FFPE RNA samples and the lower panel shows the profiling of RNA freshly extracted from human PBMCs. The Bioanalyzer profiles were obtained using Agilent RNA Nano LabChip.

Figure Legend Snippet: Bioanalyzer profiles of FFPE human peritumoral tissue total RNA. The total RNA was extracted from FFPE peritumoral tissues using Qiagen RNA extraction kit for FFPE samples or using Qiagen RNeasy Mini kit for human peripheral blood mononuclear cells (PBMCs). The upper panel shows the Bioanalyzer results of two FFPE RNA samples and the lower panel shows the profiling of RNA freshly extracted from human PBMCs. The Bioanalyzer profiles were obtained using Agilent RNA Nano LabChip.

Techniques Used: Formalin-fixed Paraffin-Embedded, RNA Extraction

17) Product Images from "Transcriptomic Profiling of Human Peritumoral Neocortex Tissues Revealed Genes Possibly Involved in Tumor-Induced Epilepsy"

Article Title: Transcriptomic Profiling of Human Peritumoral Neocortex Tissues Revealed Genes Possibly Involved in Tumor-Induced Epilepsy

Journal: PLoS ONE

doi: 10.1371/journal.pone.0056077

Techniques Used: Formalin-fixed Paraffin-Embedded, RNA Extraction

18) Product Images from "Identification of a novel monocytic phenotype in Classic Hodgkin Lymphoma tumor microenvironment"

Article Title: Identification of a novel monocytic phenotype in Classic Hodgkin Lymphoma tumor microenvironment

Journal: PLoS ONE

doi: 10.1371/journal.pone.0224621

CD163, CD14 and SR-A expression in nodal CHL, lung and liver assessed by qRT-PCR. Total RNA was prepared from FFPE sections of nodal NSCHL (n = 2) and MCCHL (n = 2), from tissue (lung and liver), and differentiated THP-1 macrophages (positive control). After reverse transcription, qRT-PCR was used to quantify expression of CD68, CD163, and SR-A. Relative expression of CD163 and SR-A in macrophages was calculated by normalizing their Ct values to that of CD68 in the same sample (ΔCt), and then to their expression in THP-1 macrophages (ΔΔCt). CD163 expression was readily detected in nodal CHL and lung/liver (tissue), and to a lesser extent in NSCHL and MCCHL. SR-A expression was readily detected in tissue, but was only detected at very low levels in NSCHL and not detected (ND) in MCCHL.

Figure Legend Snippet: CD163, CD14 and SR-A expression in nodal CHL, lung and liver assessed by qRT-PCR. Total RNA was prepared from FFPE sections of nodal NSCHL (n = 2) and MCCHL (n = 2), from tissue (lung and liver), and differentiated THP-1 macrophages (positive control). After reverse transcription, qRT-PCR was used to quantify expression of CD68, CD163, and SR-A. Relative expression of CD163 and SR-A in macrophages was calculated by normalizing their Ct values to that of CD68 in the same sample (ΔCt), and then to their expression in THP-1 macrophages (ΔΔCt). CD163 expression was readily detected in nodal CHL and lung/liver (tissue), and to a lesser extent in NSCHL and MCCHL. SR-A expression was readily detected in tissue, but was only detected at very low levels in NSCHL and not detected (ND) in MCCHL.

Techniques Used: Expressing, Quantitative RT-PCR, Formalin-fixed Paraffin-Embedded, Positive Control

19) Product Images from "Upregulation of P2Y2R, Active uPA, and PAI-1 Are Essential Components of Hantavirus Cardiopulmonary Syndrome"

Article Title: Upregulation of P2Y2R, Active uPA, and PAI-1 Are Essential Components of Hantavirus Cardiopulmonary Syndrome

Journal: Frontiers in Cellular and Infection Microbiology

doi: 10.3389/fcimb.2018.00169

Techniques Used: Expressing, Formalin-fixed Paraffin-Embedded, TaqMan Assay, Plasmid Preparation

20) Product Images from "Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies"

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

Journal: BMC Medical Research Methodology

doi: 10.1186/s12874-018-0628-1

Techniques Used: RNA Extraction, Formalin-fixed Paraffin-Embedded

21) Product Images from "Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies"

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

Journal: BMC Medical Research Methodology

doi: 10.1186/s12874-018-0628-1

Techniques Used: RNA Extraction, Formalin-fixed Paraffin-Embedded

22) Product Images from "Protein arginine methyltransferase 5 promotes bladder cancer growth through inhibiting NF-kB dependent apoptosis"

Article Title: Protein arginine methyltransferase 5 promotes bladder cancer growth through inhibiting NF-kB dependent apoptosis

Journal: EXCLI Journal

doi: 10.17179/excli2018-1719

Expression level of PRMT5 is higher in bladder cancer cell lines and primary tissues. A. Western blot of PRMT5 protein in bladder cancer cells (T24 and UM-UC-3) and control normal bladder cell lines (HBSMC and SV-HUC-1). B. PRMT5 expression level in primary bladder cancer tissues (primary cancer 1 and primary cancer 2) and control normal bladder cancer adjacent tissue (normal). C. The relative expression of PRMT5 in 10 pairs of FFPE samples from bladder cancer patients and healthy controls was analyzed by qRT-PCR. ** P

Figure Legend Snippet: Expression level of PRMT5 is higher in bladder cancer cell lines and primary tissues. A. Western blot of PRMT5 protein in bladder cancer cells (T24 and UM-UC-3) and control normal bladder cell lines (HBSMC and SV-HUC-1). B. PRMT5 expression level in primary bladder cancer tissues (primary cancer 1 and primary cancer 2) and control normal bladder cancer adjacent tissue (normal). C. The relative expression of PRMT5 in 10 pairs of FFPE samples from bladder cancer patients and healthy controls was analyzed by qRT-PCR. ** P

Techniques Used: Expressing, Western Blot, Formalin-fixed Paraffin-Embedded, Quantitative RT-PCR

23) Product Images from "Transcriptomic Profiling of Human Peritumoral Neocortex Tissues Revealed Genes Possibly Involved in Tumor-Induced Epilepsy"

Article Title: Transcriptomic Profiling of Human Peritumoral Neocortex Tissues Revealed Genes Possibly Involved in Tumor-Induced Epilepsy

Journal: PLoS ONE

doi: 10.1371/journal.pone.0056077

Techniques Used: Formalin-fixed Paraffin-Embedded, RNA Extraction

24) Product Images from "Evaluation and Validation of Total RNA Extraction Methods for MicroRNA Expression Analyses in Formalin-Fixed, Paraffin-Embedded Tissues"

Article Title: Evaluation and Validation of Total RNA Extraction Methods for MicroRNA Expression Analyses in Formalin-Fixed, Paraffin-Embedded Tissues

Journal:

doi: 10.2353/jmoldx.2008.070153

Techniques Used: Quantitative RT-PCR, Isolation, Formalin-fixed Paraffin-Embedded

25) Product Images from "Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies"

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

Journal: BMC Medical Research Methodology

doi: 10.1186/s12874-018-0628-1

Techniques Used: RNA Extraction, Formalin-fixed Paraffin-Embedded

26) Product Images from "Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies"

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

Journal: BMC Medical Research Methodology

doi: 10.1186/s12874-018-0628-1

Techniques Used: RNA Extraction, Formalin-fixed Paraffin-Embedded

27) Product Images from "Nucleic acid extraction from formalin-fixed paraffin-embedded cancer cell line samples: a trade off between quantity and quality?"

Article Title: Nucleic acid extraction from formalin-fixed paraffin-embedded cancer cell line samples: a trade off between quantity and quality?

Journal: BMC Clinical Pathology

doi: 10.1186/s12907-016-0039-3

Techniques Used: Concentration Assay, Formalin-fixed Paraffin-Embedded, Isolation, Purification

28) Product Images from "A high-throughput method to detect RNA profiling by integration of RT-MLPA with next generation sequencing technology"

Article Title: A high-throughput method to detect RNA profiling by integration of RT-MLPA with next generation sequencing technology

Journal: Oncotarget

doi: 10.18632/oncotarget.17551

The RNA expression levels of 16 cancer-associated genes in patient #04 and #05 detected by both RT-MLPSeq and RT-qPCT methods The RNA levels of 16 genes, including Ki67, STK15, Survivin, CCNB1, MYBL2, GRB7, HER2, MMP11, CTSL2, ER, PGR, BCL2, SCUBE2, GSTM1, CD68, BAG1 in FFPE sections were normalized to the average Ct value of 5 reference genes (ACTB, GAPDH, RPLPO, GUSB, TFRC).

Figure Legend Snippet: The RNA expression levels of 16 cancer-associated genes in patient #04 and #05 detected by both RT-MLPSeq and RT-qPCT methods The RNA levels of 16 genes, including Ki67, STK15, Survivin, CCNB1, MYBL2, GRB7, HER2, MMP11, CTSL2, ER, PGR, BCL2, SCUBE2, GSTM1, CD68, BAG1 in FFPE sections were normalized to the average Ct value of 5 reference genes (ACTB, GAPDH, RPLPO, GUSB, TFRC).

Techniques Used: RNA Expression, Formalin-fixed Paraffin-Embedded

Recurrence scores of 11 breast cancer patients calculated by RT-qPCR and RT-MLPSeq methods RNA from FFPE sections of 11 breast cancer patients were extracted and reverse-transcribed into cDNA. Then cDNA from the same patients were separated to determine the relative expression levels of 21 gene by RT-qPCR and MLPSeq. Recurrence scores were finally calculated according to the algorithm presented by Paik et al.

Figure Legend Snippet: Recurrence scores of 11 breast cancer patients calculated by RT-qPCR and RT-MLPSeq methods RNA from FFPE sections of 11 breast cancer patients were extracted and reverse-transcribed into cDNA. Then cDNA from the same patients were separated to determine the relative expression levels of 21 gene by RT-qPCR and MLPSeq. Recurrence scores were finally calculated according to the algorithm presented by Paik et al.

Techniques Used: Quantitative RT-PCR, Formalin-fixed Paraffin-Embedded, Expressing

29) Product Images from "Bromodomain Protein BRD4 Is a Transcriptional Repressor of Autophagy and Lysosomal Function"

Article Title: Bromodomain Protein BRD4 Is a Transcriptional Repressor of Autophagy and Lysosomal Function

Journal: Molecular Cell

doi: 10.1016/j.molcel.2017.04.027

Techniques Used: Expressing, Transfection, RNA Sequencing Assay, Quantitative RT-PCR

Techniques Used: Expressing, Transfection, Luciferase, Western Blot, Staining, Immunohistochemistry, Transgenic Assay, Mouse Assay, shRNA, Stable Transfection

30) Product Images from "CDK1 and CCNB1 as potential diagnostic markers of rhabdomyosarcoma: validation following bioinformatics analysis"

Article Title: CDK1 and CCNB1 as potential diagnostic markers of rhabdomyosarcoma: validation following bioinformatics analysis

Journal: BMC Medical Genomics

doi: 10.1186/s12920-019-0645-x

Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing

31) Product Images from "Phosphorylated TDP-43 (pTDP-43) aggregates in the axial skeletal muscle of patients with sporadic and familial amyotrophic lateral sclerosis"

Article Title: Phosphorylated TDP-43 (pTDP-43) aggregates in the axial skeletal muscle of patients with sporadic and familial amyotrophic lateral sclerosis

Journal: Acta Neuropathologica Communications

doi: 10.1186/s40478-018-0528-y

qPCR analysis was performed in 5 ALS muscle samples (“ALS1–5”) and 3 IBM muscle samples (“IBM1–3”), all containing p62 and pTDP-43 inclusions by immunohistochemical studies (see Results ). Three non-ALS, non-IBM samples with mild neurogenic atrophy in the muscle biopsy were also studied (“MSC1–3”). SQSTM1 and TARDBP were analyzed relative to the expression of two housekeeping genes ( 18 s , GAPDH ) and data shown are combined from two 96-well plates (4 replicates per sample and primer). There was a significant increase in relative normalized gene expression for both ALS and IBM samples ( a ), relative to controls (right) and this was most significant for SQSTM1 expression in ALS samples ( > 4-fold expression relative to controls, P

Figure Legend Snippet: qPCR analysis was performed in 5 ALS muscle samples (“ALS1–5”) and 3 IBM muscle samples (“IBM1–3”), all containing p62 and pTDP-43 inclusions by immunohistochemical studies (see Results ). Three non-ALS, non-IBM samples with mild neurogenic atrophy in the muscle biopsy were also studied (“MSC1–3”). SQSTM1 and TARDBP were analyzed relative to the expression of two housekeeping genes ( 18 s , GAPDH ) and data shown are combined from two 96-well plates (4 replicates per sample and primer). There was a significant increase in relative normalized gene expression for both ALS and IBM samples ( a ), relative to controls (right) and this was most significant for SQSTM1 expression in ALS samples ( > 4-fold expression relative to controls, P

Techniques Used: Real-time Polymerase Chain Reaction, Immunohistochemistry, Expressing

32) Product Images from "Quality Control of RNA Preservation and Extraction from Paraffin-Embedded Tissue: Implications for RT-PCR and Microarray Analysis"

Article Title: Quality Control of RNA Preservation and Extraction from Paraffin-Embedded Tissue: Implications for RT-PCR and Microarray Analysis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0070714

Fixation has strong influence on RNA fragment size and impairs qRT-PCR results. Electropherograms show different RNA molecule size distribution in cryopreserved and FFPE/TFPE liver tissue samples. (A) In contrast to CRYO, ribosomal RNA peaks are absent and fragmentation of RNA has occurred in all fixed tissues. (B) RNA cleanup by Qiagen RNeasy Kit removes small (

Figure Legend Snippet: Fixation has strong influence on RNA fragment size and impairs qRT-PCR results. Electropherograms show different RNA molecule size distribution in cryopreserved and FFPE/TFPE liver tissue samples. (A) In contrast to CRYO, ribosomal RNA peaks are absent and fragmentation of RNA has occurred in all fixed tissues. (B) RNA cleanup by Qiagen RNeasy Kit removes small (

Techniques Used: Quantitative RT-PCR, Formalin-fixed Paraffin-Embedded

33) Product Images from "Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies"

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

Journal: BMC Medical Research Methodology

doi: 10.1186/s12874-018-0628-1

Techniques Used: RNA Extraction, Formalin-fixed Paraffin-Embedded

34) Product Images from "Reduced expression levels of the death-associated protein kinase and E-cadherin are correlated with the development of esophageal squamous cell carcinoma"

Article Title: Reduced expression levels of the death-associated protein kinase and E-cadherin are correlated with the development of esophageal squamous cell carcinoma

Journal: Experimental and Therapeutic Medicine

doi: 10.3892/etm.2013.916

Quantitative RT-PCR analysis of DAPK and E-cadherin mRNA levels in ESCC and adjacent normal tissues. Total RNA was harvested from tissues using the RNeasy FFPE kit (Qiagen) according to the manufacturer’s instructions. The RT-PCR experiments were repeated at least 3 times. RNA was reverse transcribed into cDNA using random primers in a Reverse Transcription II system (Promega) according to the manufacturer’s instructions. Expression of DAPK and E-cadherin mRNAs was quantified by quantitative PCR using an ABI Prism Sequence Detection System (Applied Biosystems). Template-negative and RT-negative conditions were used as controls. Amplification of the endogenous GAPDH cDNA was monitored. Levels (mean values) of DAPK and E-cadherin transcripts in patients were calculated. RT-PCR, reverse transcription-PCR; ESCC, esophageal squamous cell carcinoma; DAPK, death-associated protein kinase.

Figure Legend Snippet: Quantitative RT-PCR analysis of DAPK and E-cadherin mRNA levels in ESCC and adjacent normal tissues. Total RNA was harvested from tissues using the RNeasy FFPE kit (Qiagen) according to the manufacturer’s instructions. The RT-PCR experiments were repeated at least 3 times. RNA was reverse transcribed into cDNA using random primers in a Reverse Transcription II system (Promega) according to the manufacturer’s instructions. Expression of DAPK and E-cadherin mRNAs was quantified by quantitative PCR using an ABI Prism Sequence Detection System (Applied Biosystems). Template-negative and RT-negative conditions were used as controls. Amplification of the endogenous GAPDH cDNA was monitored. Levels (mean values) of DAPK and E-cadherin transcripts in patients were calculated. RT-PCR, reverse transcription-PCR; ESCC, esophageal squamous cell carcinoma; DAPK, death-associated protein kinase.

Techniques Used: Quantitative RT-PCR, Formalin-fixed Paraffin-Embedded, Reverse Transcription Polymerase Chain Reaction, Expressing, Real-time Polymerase Chain Reaction, Sequencing, Amplification, Polymerase Chain Reaction

35) Product Images from "Nucleic acid extraction from formalin-fixed paraffin-embedded cancer cell line samples: a trade off between quantity and quality?"

Article Title: Nucleic acid extraction from formalin-fixed paraffin-embedded cancer cell line samples: a trade off between quantity and quality?

Journal: BMC Clinical Pathology

doi: 10.1186/s12907-016-0039-3

Techniques Used: Concentration Assay, Formalin-fixed Paraffin-Embedded, Isolation, Purification

36) Product Images from "Embryonic Stem Cell-Like Subpopulations in Venous Malformation"

Article Title: Embryonic Stem Cell-Like Subpopulations in Venous Malformation

Journal: Frontiers in Medicine

doi: 10.3389/fmed.2017.00162

Techniques Used: Immunohistochemistry, Staining, Expressing

Techniques Used: Staining

Techniques Used: Immunohistochemistry, Staining, Expressing

Techniques Used: Expressing, Quantitative RT-PCR

37) Product Images from "Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies"

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

Journal: BMC Medical Research Methodology

doi: 10.1186/s12874-018-0628-1

Techniques Used: RNA Extraction, Formalin-fixed Paraffin-Embedded

38) Product Images from "Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies"

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

Journal: BMC Medical Research Methodology

doi: 10.1186/s12874-018-0628-1

Techniques Used: RNA Extraction, Formalin-fixed Paraffin-Embedded

39) Product Images from "Nucleic acid extraction from formalin-fixed paraffin-embedded cancer cell line samples: a trade off between quantity and quality?"

Article Title: Nucleic acid extraction from formalin-fixed paraffin-embedded cancer cell line samples: a trade off between quantity and quality?

Journal: BMC Clinical Pathology

doi: 10.1186/s12907-016-0039-3

Investigating the relationship between DNA concentration and the input tissue amount: Geometric mean increase in DNA concentration across 6 different FFPE blocks when combining 5 μm sections in a linear fashion and associated standard error of geomean, line of Y = X/5 represents a linear relationship a Qiagen AllPrep DNA/RNA FFPE, b Qiagen QIAmp DNA FFPE tissue, c Arcturus PicoPure DNA extraction kit, d Maxwell 16 FFPE Tissue LEV DNA Purification Kit

Figure Legend Snippet: Investigating the relationship between DNA concentration and the input tissue amount: Geometric mean increase in DNA concentration across 6 different FFPE blocks when combining 5 μm sections in a linear fashion and associated standard error of geomean, line of Y = X/5 represents a linear relationship a Qiagen AllPrep DNA/RNA FFPE, b Qiagen QIAmp DNA FFPE tissue, c Arcturus PicoPure DNA extraction kit, d Maxwell 16 FFPE Tissue LEV DNA Purification Kit

Techniques Used: Concentration Assay, Formalin-fixed Paraffin-Embedded, DNA Extraction, DNA Purification

Comparison of the Agilent RNA assay and our in house PCR assay for assessing RNA quality: a Agilent RNA assay, graph shows Geometric mean RIN values and associated standard error of mean for RNA samples extracted using four different RNA extraction kits from FFPE blocks

Figure Legend Snippet: Comparison of the Agilent RNA assay and our in house PCR assay for assessing RNA quality: a Agilent RNA assay, graph shows Geometric mean RIN values and associated standard error of mean for RNA samples extracted using four different RNA extraction kits from FFPE blocks

Techniques Used: Polymerase Chain Reaction, RNA Extraction, Formalin-fixed Paraffin-Embedded

Techniques Used: Concentration Assay, Formalin-fixed Paraffin-Embedded, Isolation, Purification

40) Product Images from "Nucleic acid extraction from formalin-fixed paraffin-embedded cancer cell line samples: a trade off between quantity and quality?"

Article Title: Nucleic acid extraction from formalin-fixed paraffin-embedded cancer cell line samples: a trade off between quantity and quality?

Journal: BMC Clinical Pathology

doi: 10.1186/s12907-016-0039-3

Techniques Used: Concentration Assay, Formalin-fixed Paraffin-Embedded, Isolation, Purification

DNA Extraction:

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies
Article Snippet: .. The macro-dissected tumor area from one tissue section was used for RNA isolation using the RNeasy® FFPE kit (Qiagen, Hilden, Germany) and the tumor area from the second section was used for DNA isolation using the QIAamp® DNA FFPE Tissue kit (Qiagen, Hilden, Germany), following the manufacturers’ instructions. .. Quantity and quality measurements The quantity and purity (A260/A280) of the extracted DNA and RNA was measured using the NanoDrop ND-2000 Spectrophotometer (Thermo Scientific, Waltham, MA, USA).

Isolation:

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies
Article Snippet: .. Assessment of nucleic acids extraction kits from FFPE biopsies with normal histology The study material, consisting of 30 FFPE prostate biopsies with normal histology, was randomly divided into three different groups; 1) ten biopsies for comparison between the High Pure FFPE RNA Micro Kit (Roche Diagnostics, West Sussex, UK) and the RNeasy® FFPE kit (Qiagen, Hilden, Germany), 2) ten biopsies for comparison between the High Pure DNA FFPET Isolation Kit (Roche Diagnostics, West Sussex, UK) and the QIAamp® DNA FFPE Tissue kit (Qiagen, Hilden, Germany), and 3) ten biopsies for comparison of the best performing DNA and RNA kits from the two previous steps to the AllPrep® DNA/RNA FFPE kit (Qiagen, Hilden, Germany). .. In order to minimize the risk of variation due to different operators, the same operator performed all extractions with one kit (e.g. all extractions using the RNeasy® FFPE kit).

Formalin-fixed Paraffin-Embedded:

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies
Article Snippet: .. Based on the results from the assessment of nucleic acid extraction kits, DNA and RNA were extracted from two serial sections of the biopsies using the QIAamp® DNA FFPE Tissue Kit and RNeasy® FFPE kit. ..

Article Title: Upregulation of P2Y2R, Active uPA, and PAI-1 Are Essential Components of Hantavirus Cardiopulmonary Syndrome
Article Snippet: .. To measure the P2Y2 R expression levels in the tissue segments, RNA was extracted from 10 μm sections of the FFPE blocks using the Qiagen RNeasy FFPE kit (cat# 73504) and the instructions therein. .. The extracted RNA was quantified with a Thermo Fisher Scientific Nanodrop spectrophotometer.

Article Title: Evaluation and Validation of Total RNA Extraction Methods for MicroRNA Expression Analyses in Formalin-Fixed, Paraffin-Embedded Tissues
Article Snippet: .. In the case of the RNeasy FFPE Kit, a supplementary protocol for “co-purification” of total RNA and miRNA from FFPE tissue sections using the RNeasy FFPE Kit was used. .. Total RNA from a subset of FFPE blocks, including breast cancer, lung cancer, lung NAT, prostate NAT, spleen, and liver NAT was isolated using RecoverAll and RNeasy kits.

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies
Article Snippet: .. Figure a-c shows a comparison of RNA yields, purity (A260/A280) and RIN-values obtained from samples extracted with the High Pure FFPE RNA Micro kit and the RNeasy® FFPE kit, respectively. ..

Article Title: Nucleic acid extraction from formalin-fixed paraffin-embedded cancer cell line samples: a trade off between quantity and quality?
Article Snippet: .. The RNeasy FFPE kit gave extraction efficiencies of 86% when processing up to 20 μm of tissue, above this the efficiencies were much less. .. We felt these small losses in yield (~14%) were an acceptable compromise for the increase in concentration achieved through combining tissue sections (Fig. ).

Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies
Article Snippet: .. In order to minimize the risk of variation due to different operators, the same operator performed all extractions with one kit (e.g. all extractions using the RNeasy® FFPE kit). .. In the comparison between a) the High Pure FFPE RNA Micro kit and the RNeasy® FFPE kit and b) the High Pure FFPET DNA Isolation kit to the QIAamp® DNA FFPE Tissue kit, three serial sections of 10 μm were cut from ten biopsies with normal histology.

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