rneasy plus kit  (Qiagen)

 
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    Name:
    RNeasy Plus Mini Kit
    Description:
    For phenol free total RNA extraction from cells tissues with removal of genomic DNA contamination Kit contents Qiagen RNeasy Plus Mini Kit 50 preps 30mg Sample 30 to 50L Elution Volume Tissue Cells Sample Total RNA Purification Silica Technology Spin Column Format 25 min Time Run Ideal for PCR Northern Dot and Slot Blotting Array Analysis Sequencing For Purification of Up to 100g Total RNA from Cells tissues Using gDNA Eliminator Columns Includes RNeasy Mini Spin Columns gDNA Eliminator Spin Columns Collection Tubes RNase free Water and Buffers Benefits Efficient gDNA removal with unique gDNA Eliminator columns no need for DNase High quality total RNA in minutes using fast and simple extraction protocols Phenol free RNA isolation Ideal for sensitive applications such as real time RT PCR and RNA Seq
    Catalog Number:
    74134
    Price:
    360
    Category:
    RNeasy Plus Micro and Mini Kits
    Buy from Supplier


    Structured Review

    Qiagen rneasy plus kit
    RNeasy Plus Mini Kit
    For phenol free total RNA extraction from cells tissues with removal of genomic DNA contamination Kit contents Qiagen RNeasy Plus Mini Kit 50 preps 30mg Sample 30 to 50L Elution Volume Tissue Cells Sample Total RNA Purification Silica Technology Spin Column Format 25 min Time Run Ideal for PCR Northern Dot and Slot Blotting Array Analysis Sequencing For Purification of Up to 100g Total RNA from Cells tissues Using gDNA Eliminator Columns Includes RNeasy Mini Spin Columns gDNA Eliminator Spin Columns Collection Tubes RNase free Water and Buffers Benefits Efficient gDNA removal with unique gDNA Eliminator columns no need for DNase High quality total RNA in minutes using fast and simple extraction protocols Phenol free RNA isolation Ideal for sensitive applications such as real time RT PCR and RNA Seq
    https://www.bioz.com/result/rneasy plus kit/product/Qiagen
    Average 99 stars, based on 727 article reviews
    Price from $9.99 to $1999.99
    rneasy plus kit - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Set2 Methyltransferase Facilitates DNA Replication and Promotes Genotoxic Stress Responses through MBF-Dependent Transcription"

    Article Title: Set2 Methyltransferase Facilitates DNA Replication and Promotes Genotoxic Stress Responses through MBF-Dependent Transcription

    Journal: Cell Reports

    doi: 10.1016/j.celrep.2017.08.058

    Replication Factors Are Misregulated in set2 Δ Cells (A) Transcript levels of cdc18 + , cdc22 + , and cdt1 + were established in exponentially growing wild-type and set2 Δ cells. RNA was extracted with Qiagen RNeasy kit and relative transcript levels of cdc18 + , cdc22 + , and cdt1 + were established by RT-qPCR. Error bars represent SD of three biological repeats. The asterisk ( ∗ ) represents significant difference compared with wild-type and set2 Δ. (B) Cdc18, Cdt1, or Cdc22 protein levels of exponentially growing wild-type and set2 Δ cells. Cell extracts were prepared from vegetative cells and processed for western blot. Immunoblots of total cell lysates were probed with PAP or GFP antibody. α-Tubulin is shown as a loading control. (C) cdc18-TAP or cdc18-TAP set2 Δ cells were arrested in G1 by nitrogen starvation and released, and samples were taken at time points indicated and subjected to FACS analysis. (D) In parallel, cdc18-TAP and cdc18-TAP set2 Δ cells were processed for western blotting at indicated times. (E) cdt1-TAP or cdt1-TAP set2 Δ cells were arrested in G1 by nitrogen starvation, released, and samples taken at time points indicated and subjected to FACS analysis. (F) In parallel, cdt1-TAP and cdt1-TAP set2 Δ cells were processed for western blotting at indicated times. (G) A similar experiment to that described in (C) was carried out using cdc22-CFP or cdc22-CFP set2 Δ cells. (H) A similar experiment to that described in (D) was carried out using cdc22-CFP or cdc22-CFP set2 Δ cells. Immunoblots of total cell extracts were probed with GFP antibody. α-Tubulin is shown as a loading control.
    Figure Legend Snippet: Replication Factors Are Misregulated in set2 Δ Cells (A) Transcript levels of cdc18 + , cdc22 + , and cdt1 + were established in exponentially growing wild-type and set2 Δ cells. RNA was extracted with Qiagen RNeasy kit and relative transcript levels of cdc18 + , cdc22 + , and cdt1 + were established by RT-qPCR. Error bars represent SD of three biological repeats. The asterisk ( ∗ ) represents significant difference compared with wild-type and set2 Δ. (B) Cdc18, Cdt1, or Cdc22 protein levels of exponentially growing wild-type and set2 Δ cells. Cell extracts were prepared from vegetative cells and processed for western blot. Immunoblots of total cell lysates were probed with PAP or GFP antibody. α-Tubulin is shown as a loading control. (C) cdc18-TAP or cdc18-TAP set2 Δ cells were arrested in G1 by nitrogen starvation and released, and samples were taken at time points indicated and subjected to FACS analysis. (D) In parallel, cdc18-TAP and cdc18-TAP set2 Δ cells were processed for western blotting at indicated times. (E) cdt1-TAP or cdt1-TAP set2 Δ cells were arrested in G1 by nitrogen starvation, released, and samples taken at time points indicated and subjected to FACS analysis. (F) In parallel, cdt1-TAP and cdt1-TAP set2 Δ cells were processed for western blotting at indicated times. (G) A similar experiment to that described in (C) was carried out using cdc22-CFP or cdc22-CFP set2 Δ cells. (H) A similar experiment to that described in (D) was carried out using cdc22-CFP or cdc22-CFP set2 Δ cells. Immunoblots of total cell extracts were probed with GFP antibody. α-Tubulin is shown as a loading control.

    Techniques Used: Quantitative RT-PCR, Western Blot, FACS

    Related Articles

    Transfection:

    Article Title: The Tumor Suppressive Role of eIF3f and Its Function in Translation Inhibition and rRNA Degradation
    Article Snippet: .. Cells were harvested at 4, 8 and 16 h after transfection and total RNA was isolated using RNeasy kit (QIAGEN). rRNAs (2.0 µg) were separated on an agarose gel and visualized by UV light (B). ..

    Agarose Gel Electrophoresis:

    Article Title: The Tumor Suppressive Role of eIF3f and Its Function in Translation Inhibition and rRNA Degradation
    Article Snippet: .. Cells were harvested at 4, 8 and 16 h after transfection and total RNA was isolated using RNeasy kit (QIAGEN). rRNAs (2.0 µg) were separated on an agarose gel and visualized by UV light (B). ..

    Isolation:

    Article Title: Research Resource: A Genome-Wide Study Identifies Potential New Target Genes for POU1F1
    Article Snippet: .. Total RNA from cells infected with the lentiviral vectors was isolated by using the RNeasy Kit (Qiagen, Hilden, Germany), including on-column DNase I digestion according to the manufacturer's recommendations. .. The quality of isolated RNA was controlled by the Lab-on-Chip-System Bioanalyser 2100 (Agilent Technologies, Inc., Palo Alto, CA).

    Article Title: The Tumor Suppressive Role of eIF3f and Its Function in Translation Inhibition and rRNA Degradation
    Article Snippet: .. Cells were harvested at 4, 8 and 16 h after transfection and total RNA was isolated using RNeasy kit (QIAGEN). rRNAs (2.0 µg) were separated on an agarose gel and visualized by UV light (B). ..

    Article Title: The embryonic type of SPP1 transcriptional regulation is re-activated in glioblastoma
    Article Snippet: .. Real- time PCR Total RNA was isolated from glioma cells using Qiagen RNeasy kit, 1 μg was used as a template. .. Amplifications were performed in duplicates in 20 μl containing 2× SYBR PCR MasterMix (Applied Biosystems) and a set of primers (human SPP1, NANOG and POU5F1_1_SG/OCT4 primers were from QuantiTect Primer Assays: QT01008798, QT01844808 and QT00210840, respectively, Qiagen).

    Article Title: Mechanisms by which Porphyromonas gingivalis evades innate immunity
    Article Snippet: .. Quantitative RT-PCR BMDCs were cultured in a 24-well plate for 6 h with 108 bacteria/well and RNA was isolated from the cells by RNeasy kit (Qiagen). .. RNA was treated to remove DNA contamination (Invitrogen) and the cDNA was synthesized using random primers, and reverse transcribed using Superscript III (Invitrogen).

    Preserving:

    Article Title: High-quality RNA extraction from the sea urchin Paracentrotus lividus embryos
    Article Snippet: .. Asai et al. [ ] compared three commonly-used methods: TRIzol® , Aurum Total RNA Mini Kit and Qiagen RNeasy Micro Kit, in combination with preservation reagents TRIzol® or RNAlate r® , to obtain high-quality and large quantities of RNA from the copepod Calanus helgolandicus . .. Their results confirmed that the preservation of copepods in RNAlate r® and the extraction with Qiagen RNeasy Micro Kit were the optimal isolation method for high-quality and quantity of RNA for NGS studies of C . helgolandicus .

    Quantitative RT-PCR:

    Article Title: Mechanisms by which Porphyromonas gingivalis evades innate immunity
    Article Snippet: .. Quantitative RT-PCR BMDCs were cultured in a 24-well plate for 6 h with 108 bacteria/well and RNA was isolated from the cells by RNeasy kit (Qiagen). .. RNA was treated to remove DNA contamination (Invitrogen) and the cDNA was synthesized using random primers, and reverse transcribed using Superscript III (Invitrogen).

    Purification:

    Article Title: Techniques for the Isolation of High-Quality RNA from Cells Encapsulated in Chitosan Hydrogels
    Article Snippet: .. The protocol developed by Wang et al. uses a combination of mechanical disruption, extraction with CTAB buffer and purification with the RNeasy® kit. .. To test the efficacy of this approach with our specific material and cell source, the hydrogels were either (1) cryo-pulverized with a mortar and pestle into a fine powder or (2) finely minced with sharp surgical scissors and then disrupted using an ultrasonic dismembrator (Fisher Scientific Model 100) with three 5-s bursts at a setting of 4 with intervals of cooling on ice in 600 μL of CTAB buffer [2% CTAB, 2% poly(vinylpyrrolidone) (PVP), 1.4 M NaCl, 100 mM Tris-HCl, 20 mM EDTA, and 1% beta-mercaptoethanol in diethylpyrocarbonate (DEPC)-treated water] prewarmed to 65°C.

    Real-time Polymerase Chain Reaction:

    Article Title: The embryonic type of SPP1 transcriptional regulation is re-activated in glioblastoma
    Article Snippet: .. Real- time PCR Total RNA was isolated from glioma cells using Qiagen RNeasy kit, 1 μg was used as a template. .. Amplifications were performed in duplicates in 20 μl containing 2× SYBR PCR MasterMix (Applied Biosystems) and a set of primers (human SPP1, NANOG and POU5F1_1_SG/OCT4 primers were from QuantiTect Primer Assays: QT01008798, QT01844808 and QT00210840, respectively, Qiagen).

    Infection:

    Article Title: Research Resource: A Genome-Wide Study Identifies Potential New Target Genes for POU1F1
    Article Snippet: .. Total RNA from cells infected with the lentiviral vectors was isolated by using the RNeasy Kit (Qiagen, Hilden, Germany), including on-column DNase I digestion according to the manufacturer's recommendations. .. The quality of isolated RNA was controlled by the Lab-on-Chip-System Bioanalyser 2100 (Agilent Technologies, Inc., Palo Alto, CA).

    Cell Culture:

    Article Title: Mechanisms by which Porphyromonas gingivalis evades innate immunity
    Article Snippet: .. Quantitative RT-PCR BMDCs were cultured in a 24-well plate for 6 h with 108 bacteria/well and RNA was isolated from the cells by RNeasy kit (Qiagen). .. RNA was treated to remove DNA contamination (Invitrogen) and the cDNA was synthesized using random primers, and reverse transcribed using Superscript III (Invitrogen).

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  • 99
    Qiagen rneasy kit
    eIF3f induced rRNA degradation in pancreatic cells. (A) Restoration of eIF3f expression in pancreatic cancer cells decreased the rRNA level. MiaPaCa-2 cells were transfected with pcDNA3-eIF3f or pcDNA3. Relative fold changes of rRNA and eIF3f levels were quantified by real time RT-PCR and normalized to GAPDH mRNA. (B)(C) Silencing of eIF3f expression increased the rRNA level during apoptosis. Control and eIF3f-silenced HPDE cells were treated with DMSO vehicle control or etoposide (10 µM) for indicated time to induce apoptosis. Cells were harvested and total <t>RNA</t> was isolated using <t>RNeasy</t> Kit. Relative fold changes of 28S (B) and 18S (C) rRNA levels were quantified by real time RT-PCR after DNase treatment. (D) Time course of cell survival after etoposide treatment. HPDE cells were treated with DMSO or etoposide (10 µM) for indicated time. Cell survival was measured using MTT assay. Relative percentages of survived cells compared to DMSO-treated cells were shown. (E) Silencing of eIF3f expression attenuated rRNA degradation. Control and eIF3f-silenced HPDE cells were treated with actinomycin D (0.5 µg/ml) to block transcription for 3, 6 and 8 h before harvest. Total RNA was isolated, treated with DNase, and relative 28S and 18S rRNAs fold changes compared to control were quantified by real time RT-PCR and normalized to GAPDH mRNA.
    Rneasy Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 4510 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy kit/product/Qiagen
    Average 99 stars, based on 4510 article reviews
    Price from $9.99 to $1999.99
    rneasy kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Qiagen rna plus universal mini kit
    Response of Newly Repopulated Microglia to an Immune Challenge. Half of the mice from each dietary treatment (control or 21 d repopulation, labeled as “Repop” on figure) were administered either LPS (0.25 mg/kg) or an equivalent volume of PBS via IP injection (n = 5 per group). Mice were euthanized 6 h post-injection (following reversal testing in the Morris water maze), half of each brain was collected and snap-frozen for <t>RNA</t> extraction, <t>cDNA</t> was synthesized, and quantitative RT-PCR performed to determine mRNA gene expression for a variety of inflammatory-related markers. A) Images of the hippocampal region of each treatment are shown. B, C) No changes in microglial number or IBA1 + staining was observed in response to microglial repopulation or LPS challenge. D) The relative mRNA expression of the microglia repopulated brain in response to LPS for a variety of inflammatory gene markers is shown. Data were analyzed as a two-way ANOVA (control vs. repopulated and LPS vs. PBS) and data are presented as raw means ± SEM. Significance is dictated by the following symbols: control + PBS vs. control + LPS*; control + PBS vs. repopulated + PBS † ; control + LPS vs. repopulated + LPS # ; repopulated + PBS vs. repopulated + LPS ϕ (all comparisons, P
    Rna Plus Universal Mini Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rna plus universal mini kit/product/Qiagen
    Average 99 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    rna plus universal mini kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    eIF3f induced rRNA degradation in pancreatic cells. (A) Restoration of eIF3f expression in pancreatic cancer cells decreased the rRNA level. MiaPaCa-2 cells were transfected with pcDNA3-eIF3f or pcDNA3. Relative fold changes of rRNA and eIF3f levels were quantified by real time RT-PCR and normalized to GAPDH mRNA. (B)(C) Silencing of eIF3f expression increased the rRNA level during apoptosis. Control and eIF3f-silenced HPDE cells were treated with DMSO vehicle control or etoposide (10 µM) for indicated time to induce apoptosis. Cells were harvested and total RNA was isolated using RNeasy Kit. Relative fold changes of 28S (B) and 18S (C) rRNA levels were quantified by real time RT-PCR after DNase treatment. (D) Time course of cell survival after etoposide treatment. HPDE cells were treated with DMSO or etoposide (10 µM) for indicated time. Cell survival was measured using MTT assay. Relative percentages of survived cells compared to DMSO-treated cells were shown. (E) Silencing of eIF3f expression attenuated rRNA degradation. Control and eIF3f-silenced HPDE cells were treated with actinomycin D (0.5 µg/ml) to block transcription for 3, 6 and 8 h before harvest. Total RNA was isolated, treated with DNase, and relative 28S and 18S rRNAs fold changes compared to control were quantified by real time RT-PCR and normalized to GAPDH mRNA.

    Journal: PLoS ONE

    Article Title: The Tumor Suppressive Role of eIF3f and Its Function in Translation Inhibition and rRNA Degradation

    doi: 10.1371/journal.pone.0034194

    Figure Lengend Snippet: eIF3f induced rRNA degradation in pancreatic cells. (A) Restoration of eIF3f expression in pancreatic cancer cells decreased the rRNA level. MiaPaCa-2 cells were transfected with pcDNA3-eIF3f or pcDNA3. Relative fold changes of rRNA and eIF3f levels were quantified by real time RT-PCR and normalized to GAPDH mRNA. (B)(C) Silencing of eIF3f expression increased the rRNA level during apoptosis. Control and eIF3f-silenced HPDE cells were treated with DMSO vehicle control or etoposide (10 µM) for indicated time to induce apoptosis. Cells were harvested and total RNA was isolated using RNeasy Kit. Relative fold changes of 28S (B) and 18S (C) rRNA levels were quantified by real time RT-PCR after DNase treatment. (D) Time course of cell survival after etoposide treatment. HPDE cells were treated with DMSO or etoposide (10 µM) for indicated time. Cell survival was measured using MTT assay. Relative percentages of survived cells compared to DMSO-treated cells were shown. (E) Silencing of eIF3f expression attenuated rRNA degradation. Control and eIF3f-silenced HPDE cells were treated with actinomycin D (0.5 µg/ml) to block transcription for 3, 6 and 8 h before harvest. Total RNA was isolated, treated with DNase, and relative 28S and 18S rRNAs fold changes compared to control were quantified by real time RT-PCR and normalized to GAPDH mRNA.

    Article Snippet: Cells were harvested at 4, 8 and 16 h after transfection and total RNA was isolated using RNeasy kit (QIAGEN). rRNAs (2.0 µg) were separated on an agarose gel and visualized by UV light (B).

    Techniques: Expressing, Transfection, Quantitative RT-PCR, Isolation, MTT Assay, Blocking Assay

    Representative end-point RT-PCR gene expression results for (a) 18S and (b) TfR from RNA isolated using the freeze grind+CTAB+RNeasy ® (FCR) method, the mince+CTAB+RNeasy ® (MCR) method, and the lysozyme+CTAB+RNeasy ® (LCR) method. Genomic contamination was detected following agarose gel electrophoresis in all minus-RT controls. In addition, as shown in the TfR results, where the primers were designed to span an intron–exon boundary, two products were formed during the PCR, corresponding to a genomic product size of 270 bp and an mRNA product size of 62 bp.

    Journal: Tissue Engineering. Part C, Methods

    Article Title: Techniques for the Isolation of High-Quality RNA from Cells Encapsulated in Chitosan Hydrogels

    doi: 10.1089/ten.tec.2012.0693

    Figure Lengend Snippet: Representative end-point RT-PCR gene expression results for (a) 18S and (b) TfR from RNA isolated using the freeze grind+CTAB+RNeasy ® (FCR) method, the mince+CTAB+RNeasy ® (MCR) method, and the lysozyme+CTAB+RNeasy ® (LCR) method. Genomic contamination was detected following agarose gel electrophoresis in all minus-RT controls. In addition, as shown in the TfR results, where the primers were designed to span an intron–exon boundary, two products were formed during the PCR, corresponding to a genomic product size of 270 bp and an mRNA product size of 62 bp.

    Article Snippet: The protocol developed by Wang et al. uses a combination of mechanical disruption, extraction with CTAB buffer and purification with the RNeasy® kit.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Isolation, Agarose Gel Electrophoresis, Polymerase Chain Reaction

    SPP1 expression is up-regulated in glioma initiating cells ( A ) Representative examples of dot plots and histograms of Rhod 123(–) subpopulations from 5 glioma cell lines were acquired by flow cytometry. Glioma cells (1 × 10 7 ) were trypsinized and incubated with 0.1 μg/ml Rhod123 for 20 min in 37°C. After washing the cells were placed in 37°C for 90 min for Rhod123 exclusion in a dark compartment. Cells kept on ice to inhibit exclusion of Rhod123 were used as a positive control for gating in flow cytometry. Fractions of Rhod123(+) and Rhod123(–) cells were sorted using FACS Aria and a left panel shows a gating strategy. ( B ) Analysis of SPP1, OCT3/4 and NANOG gene expression in Rhod 123(–) subpopulations sorted from four human glioma cell lines. Total RNA was isolated from sorted cells using Qiagen RNeasy kit and the levels of SPP1, NANOG and OCT3/4 mRNA were determined by qPCR in Rhod 123(+) and Rhod 123(–) subpopulations; their expression in Rhod 123(+) subpopulations was taken as 1 (a red line). P values were calculated with a t-test and considered significant when *P ≤ 0.05 and * *P ≤ 0.01. ( C ) Quantification of the expression of Spp1, Oct3/4 and Nanog in Rhod 123(–) subpopulations isolated from rat C6 glioma cells versus their levels in Rhod 123(+) subpopulations taken as 1 (a red line); n = 3.

    Journal: Oncotarget

    Article Title: The embryonic type of SPP1 transcriptional regulation is re-activated in glioblastoma

    doi: 10.18632/oncotarget.14092

    Figure Lengend Snippet: SPP1 expression is up-regulated in glioma initiating cells ( A ) Representative examples of dot plots and histograms of Rhod 123(–) subpopulations from 5 glioma cell lines were acquired by flow cytometry. Glioma cells (1 × 10 7 ) were trypsinized and incubated with 0.1 μg/ml Rhod123 for 20 min in 37°C. After washing the cells were placed in 37°C for 90 min for Rhod123 exclusion in a dark compartment. Cells kept on ice to inhibit exclusion of Rhod123 were used as a positive control for gating in flow cytometry. Fractions of Rhod123(+) and Rhod123(–) cells were sorted using FACS Aria and a left panel shows a gating strategy. ( B ) Analysis of SPP1, OCT3/4 and NANOG gene expression in Rhod 123(–) subpopulations sorted from four human glioma cell lines. Total RNA was isolated from sorted cells using Qiagen RNeasy kit and the levels of SPP1, NANOG and OCT3/4 mRNA were determined by qPCR in Rhod 123(+) and Rhod 123(–) subpopulations; their expression in Rhod 123(+) subpopulations was taken as 1 (a red line). P values were calculated with a t-test and considered significant when *P ≤ 0.05 and * *P ≤ 0.01. ( C ) Quantification of the expression of Spp1, Oct3/4 and Nanog in Rhod 123(–) subpopulations isolated from rat C6 glioma cells versus their levels in Rhod 123(+) subpopulations taken as 1 (a red line); n = 3.

    Article Snippet: Real- time PCR Total RNA was isolated from glioma cells using Qiagen RNeasy kit, 1 μg was used as a template.

    Techniques: Expressing, Flow Cytometry, Cytometry, Incubation, Positive Control, FACS, Isolation, Real-time Polymerase Chain Reaction

    Response of Newly Repopulated Microglia to an Immune Challenge. Half of the mice from each dietary treatment (control or 21 d repopulation, labeled as “Repop” on figure) were administered either LPS (0.25 mg/kg) or an equivalent volume of PBS via IP injection (n = 5 per group). Mice were euthanized 6 h post-injection (following reversal testing in the Morris water maze), half of each brain was collected and snap-frozen for RNA extraction, cDNA was synthesized, and quantitative RT-PCR performed to determine mRNA gene expression for a variety of inflammatory-related markers. A) Images of the hippocampal region of each treatment are shown. B, C) No changes in microglial number or IBA1 + staining was observed in response to microglial repopulation or LPS challenge. D) The relative mRNA expression of the microglia repopulated brain in response to LPS for a variety of inflammatory gene markers is shown. Data were analyzed as a two-way ANOVA (control vs. repopulated and LPS vs. PBS) and data are presented as raw means ± SEM. Significance is dictated by the following symbols: control + PBS vs. control + LPS*; control + PBS vs. repopulated + PBS † ; control + LPS vs. repopulated + LPS # ; repopulated + PBS vs. repopulated + LPS ϕ (all comparisons, P

    Journal: PLoS ONE

    Article Title: Characterizing Newly Repopulated Microglia in the Adult Mouse: Impacts on Animal Behavior, Cell Morphology, and Neuroinflammation

    doi: 10.1371/journal.pone.0122912

    Figure Lengend Snippet: Response of Newly Repopulated Microglia to an Immune Challenge. Half of the mice from each dietary treatment (control or 21 d repopulation, labeled as “Repop” on figure) were administered either LPS (0.25 mg/kg) or an equivalent volume of PBS via IP injection (n = 5 per group). Mice were euthanized 6 h post-injection (following reversal testing in the Morris water maze), half of each brain was collected and snap-frozen for RNA extraction, cDNA was synthesized, and quantitative RT-PCR performed to determine mRNA gene expression for a variety of inflammatory-related markers. A) Images of the hippocampal region of each treatment are shown. B, C) No changes in microglial number or IBA1 + staining was observed in response to microglial repopulation or LPS challenge. D) The relative mRNA expression of the microglia repopulated brain in response to LPS for a variety of inflammatory gene markers is shown. Data were analyzed as a two-way ANOVA (control vs. repopulated and LPS vs. PBS) and data are presented as raw means ± SEM. Significance is dictated by the following symbols: control + PBS vs. control + LPS*; control + PBS vs. repopulated + PBS † ; control + LPS vs. repopulated + LPS # ; repopulated + PBS vs. repopulated + LPS ϕ (all comparisons, P

    Article Snippet: RNA was extracted (RNA Plus Universal Mini Kit, Qiagen), cDNA synthesized (iScript kit, BioRad), and quantitative RT-PCR performed using a commercially available immune panel that tests 96 genes (including pro-inflammatory cytokines, chemokines, and microglial specific markers; Taqman Array Mouse Immune Panel; Invitrogen). mRNA expression data were analyzed using the comparative threshold cycle (Ct) method [ ] and results are expressed as percent change from controls administered PBS.

    Techniques: Mouse Assay, Labeling, Injection, RNA Extraction, Synthesized, Quantitative RT-PCR, Expressing, Staining