rna  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier
Bioz Manufacturer Symbol Thermo Fisher manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99

    Structured Review

    Thermo Fisher rna
    hnRNP H and hnRNP F modulate HRAS exon 5 splicing through G-run elements within the downstream ISE. (A) Diagram of HRAS minigene reporters carrying mutations at putative hnRNP H/F binding motifs. Intron 5 nucleotide sequences targeted by ASO’s I5-1 to I5-8 are shown in orange as potential enhancers and green as potential suppressers. G-runs within this intron 5 region is underlined, and those that are within the enhancer regions and mutated are labeled in red and numbered. Mutations in these elements are indicated below, with the mutated sequences underlined and labeled in red. The known silencer element rasISS1 is boxed in navy and its putative hnRNP A1 binding motif is highlighted ( Guil et al., 2003b ). (B) Immunoblot showing the expression of endogenous hnRNP’s H and F and His-tagged ectopic proteins. GAPDH was used as the loading control. RT-PCR analyses showing the splicing changes of wild type and mutant minigene reporters after co-transfection with control (top), hnRNP H (middle), and hnRNP F (bottom) expression plasmids (wt, wild type; mt, mutant). The bar graph presents the quantification of RT-PCR results, with the mean +/- SD of three biological replicates. (C) <t>RNA</t> immunoprecipitation assayed by real-time RT-PCR of wildtype and G1G2 mutant HRAS intron 5 RNA bound by hnRNP H and hnRNP F proteins. HEK293 cells transfected with wtHRAS or mtG1G2 minigenes were immunoprecipitated using anti-epitope tag (Flag or His) antibodies, anti-endogenous protein (hnRNP H or hnRNP F) antibodies, or non-immune IgG. The immunoblot shows the recovery of hnRNP H, hnRNP F or both in the IP’s. (D) The bar graphs (right) of real-time RT-PCR quantification of recovered RNA using two sets of primers: the I5 product targets an intronic region neighboring the mutation and the E4I4 product targets the sequence immediately upstream.
    Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rna/product/Thermo Fisher
    Average 99 stars, based on 40 article reviews
    Price from $9.99 to $1999.99
    rna - by Bioz Stars, 2022-12
    99/100 stars

    Images

    1) Product Images from "The RNA binding proteins hnRNP H and F regulate splicing of a MYC dependent HRAS exon in Prostate Cancer Cells"

    Article Title: The RNA binding proteins hnRNP H and F regulate splicing of a MYC dependent HRAS exon in Prostate Cancer Cells

    Journal: bioRxiv

    doi: 10.1101/2022.11.29.518269

    hnRNP H and hnRNP F modulate HRAS exon 5 splicing through G-run elements within the downstream ISE. (A) Diagram of HRAS minigene reporters carrying mutations at putative hnRNP H/F binding motifs. Intron 5 nucleotide sequences targeted by ASO’s I5-1 to I5-8 are shown in orange as potential enhancers and green as potential suppressers. G-runs within this intron 5 region is underlined, and those that are within the enhancer regions and mutated are labeled in red and numbered. Mutations in these elements are indicated below, with the mutated sequences underlined and labeled in red. The known silencer element rasISS1 is boxed in navy and its putative hnRNP A1 binding motif is highlighted ( Guil et al., 2003b ). (B) Immunoblot showing the expression of endogenous hnRNP’s H and F and His-tagged ectopic proteins. GAPDH was used as the loading control. RT-PCR analyses showing the splicing changes of wild type and mutant minigene reporters after co-transfection with control (top), hnRNP H (middle), and hnRNP F (bottom) expression plasmids (wt, wild type; mt, mutant). The bar graph presents the quantification of RT-PCR results, with the mean +/- SD of three biological replicates. (C) RNA immunoprecipitation assayed by real-time RT-PCR of wildtype and G1G2 mutant HRAS intron 5 RNA bound by hnRNP H and hnRNP F proteins. HEK293 cells transfected with wtHRAS or mtG1G2 minigenes were immunoprecipitated using anti-epitope tag (Flag or His) antibodies, anti-endogenous protein (hnRNP H or hnRNP F) antibodies, or non-immune IgG. The immunoblot shows the recovery of hnRNP H, hnRNP F or both in the IP’s. (D) The bar graphs (right) of real-time RT-PCR quantification of recovered RNA using two sets of primers: the I5 product targets an intronic region neighboring the mutation and the E4I4 product targets the sequence immediately upstream.
    Figure Legend Snippet: hnRNP H and hnRNP F modulate HRAS exon 5 splicing through G-run elements within the downstream ISE. (A) Diagram of HRAS minigene reporters carrying mutations at putative hnRNP H/F binding motifs. Intron 5 nucleotide sequences targeted by ASO’s I5-1 to I5-8 are shown in orange as potential enhancers and green as potential suppressers. G-runs within this intron 5 region is underlined, and those that are within the enhancer regions and mutated are labeled in red and numbered. Mutations in these elements are indicated below, with the mutated sequences underlined and labeled in red. The known silencer element rasISS1 is boxed in navy and its putative hnRNP A1 binding motif is highlighted ( Guil et al., 2003b ). (B) Immunoblot showing the expression of endogenous hnRNP’s H and F and His-tagged ectopic proteins. GAPDH was used as the loading control. RT-PCR analyses showing the splicing changes of wild type and mutant minigene reporters after co-transfection with control (top), hnRNP H (middle), and hnRNP F (bottom) expression plasmids (wt, wild type; mt, mutant). The bar graph presents the quantification of RT-PCR results, with the mean +/- SD of three biological replicates. (C) RNA immunoprecipitation assayed by real-time RT-PCR of wildtype and G1G2 mutant HRAS intron 5 RNA bound by hnRNP H and hnRNP F proteins. HEK293 cells transfected with wtHRAS or mtG1G2 minigenes were immunoprecipitated using anti-epitope tag (Flag or His) antibodies, anti-endogenous protein (hnRNP H or hnRNP F) antibodies, or non-immune IgG. The immunoblot shows the recovery of hnRNP H, hnRNP F or both in the IP’s. (D) The bar graphs (right) of real-time RT-PCR quantification of recovered RNA using two sets of primers: the I5 product targets an intronic region neighboring the mutation and the E4I4 product targets the sequence immediately upstream.

    Techniques Used: Binding Assay, Allele-specific Oligonucleotide, Labeling, Expressing, Reverse Transcription Polymerase Chain Reaction, Mutagenesis, Cotransfection, Immunoprecipitation, Quantitative RT-PCR, Transfection, Sequencing

    2) Product Images from "Oxygen is a critical regulator of cellular metabolism and function in cell culture"

    Article Title: Oxygen is a critical regulator of cellular metabolism and function in cell culture

    Journal: bioRxiv

    doi: 10.1101/2022.11.29.516437

    Lowering medium volumes reduces lactate production and improves functional outcomes in other cell types and organoids (A) Extracellular medium glucose and lactate measurements after 16 h medium volume change in murine brown adipocytes (pBAT) (n = 8 biological replicates) and L6 myotubes (n = 3 biological replicates). (B) Lactate secretion in iPSC-derived hepatocytes (High = 1 mL, Low = 0.5 mL) after 24 h of medium volume change (n = 7 biological replicates). (C) Relative RNA expression of hepatocyte differentiation marker genes. Cells were cultured in either 1 mL or 0.5 mL of medium throughout differentiation, and switched to either 1 mL or 0.5 mL of medium 24 h prior to the experiment (n = 3 biological replicates). * E = significance due to end medium volume, * S = significance due to starting medium volume, * S x E = significance due to interaction between starting and end media volumes. (D) Relative CYP3A4 activity in iPSC-derived hepatocytes after 24 h of medium volume change (n = 6 biological replicates). High = 1 mL, Low = 0.5 mL. (E) Immunofluorescence of albumin in iPSC-derived hepatocytes after 24 h of medium volume change (n = 3 biological replicates). Scale bar = 350 μm. (F) Lactate secretion in cardiac organoids after 48 h of medium volume change (normalised) (n = 3-6 technical replicates from n = 3 biological replicates). High = 150 μL, Low = 50 μL. (G) Cardiac contractile force (normalised) (n = 2-17 technical replicates from n = 4 biological replicates). All data are represented as mean ± SEM. * p
    Figure Legend Snippet: Lowering medium volumes reduces lactate production and improves functional outcomes in other cell types and organoids (A) Extracellular medium glucose and lactate measurements after 16 h medium volume change in murine brown adipocytes (pBAT) (n = 8 biological replicates) and L6 myotubes (n = 3 biological replicates). (B) Lactate secretion in iPSC-derived hepatocytes (High = 1 mL, Low = 0.5 mL) after 24 h of medium volume change (n = 7 biological replicates). (C) Relative RNA expression of hepatocyte differentiation marker genes. Cells were cultured in either 1 mL or 0.5 mL of medium throughout differentiation, and switched to either 1 mL or 0.5 mL of medium 24 h prior to the experiment (n = 3 biological replicates). * E = significance due to end medium volume, * S = significance due to starting medium volume, * S x E = significance due to interaction between starting and end media volumes. (D) Relative CYP3A4 activity in iPSC-derived hepatocytes after 24 h of medium volume change (n = 6 biological replicates). High = 1 mL, Low = 0.5 mL. (E) Immunofluorescence of albumin in iPSC-derived hepatocytes after 24 h of medium volume change (n = 3 biological replicates). Scale bar = 350 μm. (F) Lactate secretion in cardiac organoids after 48 h of medium volume change (normalised) (n = 3-6 technical replicates from n = 3 biological replicates). High = 150 μL, Low = 50 μL. (G) Cardiac contractile force (normalised) (n = 2-17 technical replicates from n = 4 biological replicates). All data are represented as mean ± SEM. * p

    Techniques Used: Functional Assay, Derivative Assay, RNA Expression, Marker, Cell Culture, Activity Assay, Immunofluorescence

    Lowering medium volumes induces a widespread transcriptional response and improves adipocyte function (A) Western blot of hypoxia-inducible factor (HIF) 1α after 16 h medium volume change, with 500 μM CoCl 2 as positive control (n = 3 biological replicates). (B) Relative RNA expression of HIF1α target genes in 3T3-L1 adipocytes after 16 h medium volume change (n = 3 biological replicates). (C) Volcano plot of differentially expressed genes after 16 h medium volume change (n = 6 biological replicates). (D) Rate of leptin secretion under different medium volume conditions (n = 4 biological replicates). (E) Rate of adiponectin secretion under different medium volume conditions (n = 4 biological replicates). (F) Fluorescence intensity of plasma membrane (PM) GLUT4 upon insulin stimulation after 48 h medium volume change (n = 4 biological replicates). (G) Dose response curve of lipolytic drug, CL316,243 treatment (n = 7 biological replicates). (H) Rate of lipolysis measured by glycerol release, upon 100 nM insulin or 1 nM CL316,243 stimulation (n = 4 biological replicates). (I) KEGG pathway analyses of 3T3-L1 adipocytes after 16 h medium volume change (n = 6 biological replicates) and subcutaneous white adipose tissue (scWAT) from mice kept in 10% or 21% oxygen for 4 weeks (n = 10 biological replicates). Orange (positive NES) represents upregulated KEGG pathways in high medium (3T3-L1 adipocytes) or in 10% oxygen (mice scWAT). Blue (negative NES) represents upregulated KEGG pathways in low medium (3T3-L1 adipocytes) or in 21% oxygen (mice scWAT). NES, normalised enrichment score. Data are represented as mean ± SEM. * p
    Figure Legend Snippet: Lowering medium volumes induces a widespread transcriptional response and improves adipocyte function (A) Western blot of hypoxia-inducible factor (HIF) 1α after 16 h medium volume change, with 500 μM CoCl 2 as positive control (n = 3 biological replicates). (B) Relative RNA expression of HIF1α target genes in 3T3-L1 adipocytes after 16 h medium volume change (n = 3 biological replicates). (C) Volcano plot of differentially expressed genes after 16 h medium volume change (n = 6 biological replicates). (D) Rate of leptin secretion under different medium volume conditions (n = 4 biological replicates). (E) Rate of adiponectin secretion under different medium volume conditions (n = 4 biological replicates). (F) Fluorescence intensity of plasma membrane (PM) GLUT4 upon insulin stimulation after 48 h medium volume change (n = 4 biological replicates). (G) Dose response curve of lipolytic drug, CL316,243 treatment (n = 7 biological replicates). (H) Rate of lipolysis measured by glycerol release, upon 100 nM insulin or 1 nM CL316,243 stimulation (n = 4 biological replicates). (I) KEGG pathway analyses of 3T3-L1 adipocytes after 16 h medium volume change (n = 6 biological replicates) and subcutaneous white adipose tissue (scWAT) from mice kept in 10% or 21% oxygen for 4 weeks (n = 10 biological replicates). Orange (positive NES) represents upregulated KEGG pathways in high medium (3T3-L1 adipocytes) or in 10% oxygen (mice scWAT). Blue (negative NES) represents upregulated KEGG pathways in low medium (3T3-L1 adipocytes) or in 21% oxygen (mice scWAT). NES, normalised enrichment score. Data are represented as mean ± SEM. * p

    Techniques Used: Western Blot, Positive Control, RNA Expression, Fluorescence, Mouse Assay

    3) Product Images from "Improved bacterial single-cell RNA-seq through automated MATQ-seq and Cas9-based removal of rRNA reads"

    Article Title: Improved bacterial single-cell RNA-seq through automated MATQ-seq and Cas9-based removal of rRNA reads

    Journal: bioRxiv

    doi: 10.1101/2022.11.28.518171

    Validation of reverse transcriptases. Bioanalyzer profiles of cDNA processed with MATQ-seq using different reverse transcriptases. A) Further validation of the three RTs SS IV, Maxima H Minus and TGIRT after initial selection and subsequent buffer optimization ( Fig. 2A ). All assays were performed with a spike-in of 50 pg total RNA. B) cDNA concentrations of samples in A) measured with a Qubit fluorometer. C) Comparison of cDNA profiles obtained using SS III and SS IV (left panel) and Maxima H Minus (right panel). For SS IV and Maxima H Minus a gradient from 50, 10 and single sorted cells as input material is shown. L: ladder; NC: negative control; PC: positive control; sc: single-cell. SS III: SuperScript III; SS IV: SuperScript IV; Maxima H-: Maxima H Minus.
    Figure Legend Snippet: Validation of reverse transcriptases. Bioanalyzer profiles of cDNA processed with MATQ-seq using different reverse transcriptases. A) Further validation of the three RTs SS IV, Maxima H Minus and TGIRT after initial selection and subsequent buffer optimization ( Fig. 2A ). All assays were performed with a spike-in of 50 pg total RNA. B) cDNA concentrations of samples in A) measured with a Qubit fluorometer. C) Comparison of cDNA profiles obtained using SS III and SS IV (left panel) and Maxima H Minus (right panel). For SS IV and Maxima H Minus a gradient from 50, 10 and single sorted cells as input material is shown. L: ladder; NC: negative control; PC: positive control; sc: single-cell. SS III: SuperScript III; SS IV: SuperScript IV; Maxima H-: Maxima H Minus.

    Techniques Used: Selection, Negative Control, Positive Control

    4) Product Images from "H3K4 di- and trimethylation modulate the stability of RNA polymerase II pausing"

    Article Title: H3K4 di- and trimethylation modulate the stability of RNA polymerase II pausing

    Journal: bioRxiv

    doi: 10.1101/2022.11.28.518073

    The loss of H3K4me2 and H3K4me3 compromise gene expression (A) Metaplot showing the average Pol II occupancy genes with H3K4me3 promoter peaks as measured by ChIP-Rx in mRBBP5-dTAG cells treated with DMSO or dTAG. Pol II occupancy is represented by its largest subunit RPB1. (B and C) Two-dimensional density plot comparing the log2 fold change of Pol II ChIP-Rx signal (B) and PRO-seq signal (C) at promoters (x axis) and gene bodies (y axis) (dTAG versus DMSO) for genes with H3K4me3 promoter peaks. (D) Western blotting of mSPT5-dTAG mESCs with indicated time-course of dTAG treatment. (E and F) Metaplots showing the average level of INTS5 (E) and INTS11 (F) occupancies centered at the TSS of genes with H3K4me3 promoter peaks. (G) Boxplot showing the relative occupancies of INTS5 and INTS11 compared with Pol II in mRBBP5-dTAG mESCs treated with DMSO or dTAG. (H) M (log2 fold change, y axis) versus A (log2 average normalized read abundance, x axis) (MA) plot of spike-in normalized RNA-seq showing the change in gene expression by dTAG treatment for 24 hours in mRBBP5-dTAG cells. (I) Two-dimensional density plot comparing the log2 fold change of Pol II ChIP-Rx signal at promoters (x axis) and RNA-seq signal (y axis) (dTAG versus DMSO) for genes with H3K4me3 promoter peaks. (J and K) Boxplots showing the correlation of log2 fold change of RNA-seq singals (dTAG versus DMSO) and H3K4me3 (J) or H3K4me2 (K) (dTAG versus DMSO, grouped based on the declining extent of H3K4me3/H3K4me2 occupancy at promoters) in mRBBP5-dTAG mESCs. (L and M) Boxplots showing the correlation of log2 fold change of RNA-seq signal (dTAG versus DMSO) and H3K4me3 (L) or H3K4me2 (M) (dTAG versus DMSO, grouped based on the declining extent of H3K4me3/H3K4me2 occupancy at promoters) in mDPY30-dTAG mESCs. See also Figure S5 .
    Figure Legend Snippet: The loss of H3K4me2 and H3K4me3 compromise gene expression (A) Metaplot showing the average Pol II occupancy genes with H3K4me3 promoter peaks as measured by ChIP-Rx in mRBBP5-dTAG cells treated with DMSO or dTAG. Pol II occupancy is represented by its largest subunit RPB1. (B and C) Two-dimensional density plot comparing the log2 fold change of Pol II ChIP-Rx signal (B) and PRO-seq signal (C) at promoters (x axis) and gene bodies (y axis) (dTAG versus DMSO) for genes with H3K4me3 promoter peaks. (D) Western blotting of mSPT5-dTAG mESCs with indicated time-course of dTAG treatment. (E and F) Metaplots showing the average level of INTS5 (E) and INTS11 (F) occupancies centered at the TSS of genes with H3K4me3 promoter peaks. (G) Boxplot showing the relative occupancies of INTS5 and INTS11 compared with Pol II in mRBBP5-dTAG mESCs treated with DMSO or dTAG. (H) M (log2 fold change, y axis) versus A (log2 average normalized read abundance, x axis) (MA) plot of spike-in normalized RNA-seq showing the change in gene expression by dTAG treatment for 24 hours in mRBBP5-dTAG cells. (I) Two-dimensional density plot comparing the log2 fold change of Pol II ChIP-Rx signal at promoters (x axis) and RNA-seq signal (y axis) (dTAG versus DMSO) for genes with H3K4me3 promoter peaks. (J and K) Boxplots showing the correlation of log2 fold change of RNA-seq singals (dTAG versus DMSO) and H3K4me3 (J) or H3K4me2 (K) (dTAG versus DMSO, grouped based on the declining extent of H3K4me3/H3K4me2 occupancy at promoters) in mRBBP5-dTAG mESCs. (L and M) Boxplots showing the correlation of log2 fold change of RNA-seq signal (dTAG versus DMSO) and H3K4me3 (L) or H3K4me2 (M) (dTAG versus DMSO, grouped based on the declining extent of H3K4me3/H3K4me2 occupancy at promoters) in mDPY30-dTAG mESCs. See also Figure S5 .

    Techniques Used: Expressing, Chromatin Immunoprecipitation, Western Blot, RNA Sequencing Assay

    (A) Boxplots showing the correlation of the log2 fold change of Pol II at promoters and gene bodies (dTAG versus DMSO) for four equal groups based on the fold change of Pol II at promoters in mRBBP5-dTAG mESCs. (B) Boxplots showing the correlation of the log2 fold change of PRO-seq signal at promoters and gene bodies (dTAG versus DMSO) for four equal groups based on the fold change of PRO-seq signal at promoters in mRBBP5-dTAG mESCs. (C and D) Heatmaps of INTS5 (C) and INTS11 (D) occupancies (RPM per bp and log2 fold change) centered at the TSS of genes with H3K4me3 promoter peaks. (E) Boxplots showing the relative occupancies of INTS5 and INTS11 compared with SPT5 in mRBBP5-dTAG mESCs treated with DMSO or dTAG. (F) MA plot of spike-in normalized RNA-seq showing the gene expression changes by dTAG treatment for 24 hours in mDPY30-dTAG cells.
    Figure Legend Snippet: (A) Boxplots showing the correlation of the log2 fold change of Pol II at promoters and gene bodies (dTAG versus DMSO) for four equal groups based on the fold change of Pol II at promoters in mRBBP5-dTAG mESCs. (B) Boxplots showing the correlation of the log2 fold change of PRO-seq signal at promoters and gene bodies (dTAG versus DMSO) for four equal groups based on the fold change of PRO-seq signal at promoters in mRBBP5-dTAG mESCs. (C and D) Heatmaps of INTS5 (C) and INTS11 (D) occupancies (RPM per bp and log2 fold change) centered at the TSS of genes with H3K4me3 promoter peaks. (E) Boxplots showing the relative occupancies of INTS5 and INTS11 compared with SPT5 in mRBBP5-dTAG mESCs treated with DMSO or dTAG. (F) MA plot of spike-in normalized RNA-seq showing the gene expression changes by dTAG treatment for 24 hours in mDPY30-dTAG cells.

    Techniques Used: RNA Sequencing Assay, Expressing

    5) Product Images from "Oxygen is a critical regulator of cellular metabolism and function in cell culture"

    Article Title: Oxygen is a critical regulator of cellular metabolism and function in cell culture

    Journal: bioRxiv

    doi: 10.1101/2022.11.29.516437

    Lowering medium volumes reduces lactate production and improves functional outcomes in other cell types and organoids (A) Extracellular medium glucose and lactate measurements after 16 h medium volume change in murine brown adipocytes (pBAT) (n = 8 biological replicates) and L6 myotubes (n = 3 biological replicates). (B) Lactate secretion in iPSC-derived hepatocytes (High = 1 mL, Low = 0.5 mL) after 24 h of medium volume change (n = 7 biological replicates). (C) Relative RNA expression of hepatocyte differentiation marker genes. Cells were cultured in either 1 mL or 0.5 mL of medium throughout differentiation, and switched to either 1 mL or 0.5 mL of medium 24 h prior to the experiment (n = 3 biological replicates). * E = significance due to end medium volume, * S = significance due to starting medium volume, * S x E = significance due to interaction between starting and end media volumes. (D) Relative CYP3A4 activity in iPSC-derived hepatocytes after 24 h of medium volume change (n = 6 biological replicates). High = 1 mL, Low = 0.5 mL. (E) Immunofluorescence of albumin in iPSC-derived hepatocytes after 24 h of medium volume change (n = 3 biological replicates). Scale bar = 350 μm. (F) Lactate secretion in cardiac organoids after 48 h of medium volume change (normalised) (n = 3-6 technical replicates from n = 3 biological replicates). High = 150 μL, Low = 50 μL. (G) Cardiac contractile force (normalised) (n = 2-17 technical replicates from n = 4 biological replicates). All data are represented as mean ± SEM. * p
    Figure Legend Snippet: Lowering medium volumes reduces lactate production and improves functional outcomes in other cell types and organoids (A) Extracellular medium glucose and lactate measurements after 16 h medium volume change in murine brown adipocytes (pBAT) (n = 8 biological replicates) and L6 myotubes (n = 3 biological replicates). (B) Lactate secretion in iPSC-derived hepatocytes (High = 1 mL, Low = 0.5 mL) after 24 h of medium volume change (n = 7 biological replicates). (C) Relative RNA expression of hepatocyte differentiation marker genes. Cells were cultured in either 1 mL or 0.5 mL of medium throughout differentiation, and switched to either 1 mL or 0.5 mL of medium 24 h prior to the experiment (n = 3 biological replicates). * E = significance due to end medium volume, * S = significance due to starting medium volume, * S x E = significance due to interaction between starting and end media volumes. (D) Relative CYP3A4 activity in iPSC-derived hepatocytes after 24 h of medium volume change (n = 6 biological replicates). High = 1 mL, Low = 0.5 mL. (E) Immunofluorescence of albumin in iPSC-derived hepatocytes after 24 h of medium volume change (n = 3 biological replicates). Scale bar = 350 μm. (F) Lactate secretion in cardiac organoids after 48 h of medium volume change (normalised) (n = 3-6 technical replicates from n = 3 biological replicates). High = 150 μL, Low = 50 μL. (G) Cardiac contractile force (normalised) (n = 2-17 technical replicates from n = 4 biological replicates). All data are represented as mean ± SEM. * p

    Techniques Used: Functional Assay, Derivative Assay, RNA Expression, Marker, Cell Culture, Activity Assay, Immunofluorescence

    Lowering medium volumes induces a widespread transcriptional response and improves adipocyte function (A) Western blot of hypoxia-inducible factor (HIF) 1α after 16 h medium volume change, with 500 μM CoCl 2 as positive control (n = 3 biological replicates). (B) Relative RNA expression of HIF1α target genes in 3T3-L1 adipocytes after 16 h medium volume change (n = 3 biological replicates). (C) Volcano plot of differentially expressed genes after 16 h medium volume change (n = 6 biological replicates). (D) Rate of leptin secretion under different medium volume conditions (n = 4 biological replicates). (E) Rate of adiponectin secretion under different medium volume conditions (n = 4 biological replicates). (F) Fluorescence intensity of plasma membrane (PM) GLUT4 upon insulin stimulation after 48 h medium volume change (n = 4 biological replicates). (G) Dose response curve of lipolytic drug, CL316,243 treatment (n = 7 biological replicates). (H) Rate of lipolysis measured by glycerol release, upon 100 nM insulin or 1 nM CL316,243 stimulation (n = 4 biological replicates). (I) KEGG pathway analyses of 3T3-L1 adipocytes after 16 h medium volume change (n = 6 biological replicates) and subcutaneous white adipose tissue (scWAT) from mice kept in 10% or 21% oxygen for 4 weeks (n = 10 biological replicates). Orange (positive NES) represents upregulated KEGG pathways in high medium (3T3-L1 adipocytes) or in 10% oxygen (mice scWAT). Blue (negative NES) represents upregulated KEGG pathways in low medium (3T3-L1 adipocytes) or in 21% oxygen (mice scWAT). NES, normalised enrichment score. Data are represented as mean ± SEM. * p
    Figure Legend Snippet: Lowering medium volumes induces a widespread transcriptional response and improves adipocyte function (A) Western blot of hypoxia-inducible factor (HIF) 1α after 16 h medium volume change, with 500 μM CoCl 2 as positive control (n = 3 biological replicates). (B) Relative RNA expression of HIF1α target genes in 3T3-L1 adipocytes after 16 h medium volume change (n = 3 biological replicates). (C) Volcano plot of differentially expressed genes after 16 h medium volume change (n = 6 biological replicates). (D) Rate of leptin secretion under different medium volume conditions (n = 4 biological replicates). (E) Rate of adiponectin secretion under different medium volume conditions (n = 4 biological replicates). (F) Fluorescence intensity of plasma membrane (PM) GLUT4 upon insulin stimulation after 48 h medium volume change (n = 4 biological replicates). (G) Dose response curve of lipolytic drug, CL316,243 treatment (n = 7 biological replicates). (H) Rate of lipolysis measured by glycerol release, upon 100 nM insulin or 1 nM CL316,243 stimulation (n = 4 biological replicates). (I) KEGG pathway analyses of 3T3-L1 adipocytes after 16 h medium volume change (n = 6 biological replicates) and subcutaneous white adipose tissue (scWAT) from mice kept in 10% or 21% oxygen for 4 weeks (n = 10 biological replicates). Orange (positive NES) represents upregulated KEGG pathways in high medium (3T3-L1 adipocytes) or in 10% oxygen (mice scWAT). Blue (negative NES) represents upregulated KEGG pathways in low medium (3T3-L1 adipocytes) or in 21% oxygen (mice scWAT). NES, normalised enrichment score. Data are represented as mean ± SEM. * p

    Techniques Used: Western Blot, Positive Control, RNA Expression, Fluorescence, Mouse Assay

    6) Product Images from "Single-Cell Epitope-Transcriptomics Reveal Lung Stromal and Immune Cell Response Kinetics to Nanoparticle-delivered RIG-I and TLR4 Agonists"

    Article Title: Single-Cell Epitope-Transcriptomics Reveal Lung Stromal and Immune Cell Response Kinetics to Nanoparticle-delivered RIG-I and TLR4 Agonists

    Journal: bioRxiv

    doi: 10.1101/2022.11.29.518316

    Synthetic, experimental, and analytical overview for PUUC and MPLA+PUUC nanoparticle stimulation at early time points in the murine lung. A) Schematic of nanoparticle (NP) synthesis. PLGA NPs with or without mono-phosphoryl lipid A (MPLA) were first synthesized by double-emulsion antisolvent evaporation, followed by covalent modification with branched polyethyleneimine (PEI) with EDC/sulfo-NHS to make positively-charged PLGA-PEI NPs. Negatively charged PUUC RNA was then electrostatically loaded onto the particles to make either PUUC or MPLA+PUUC NPs. B) Schematic of experimental and analytical overview. 4 mg of PUUC and MPLA+PUUC NPs were combined with 60 μL saline and were administered into the bilateral nares of three mice per group. Doses of 20 ug PUUC and 24 ug MPLA were used per mouse. Lungs from these groups and naïve, untreated mice were harvested, processed into single-cell suspensions, and were pooled groupwise at 4 and 24 hours. Suspensions were mixed with DNA-barcoded antibodies (antibody-dependent tags; ADTs) and were processed according to 10X Chromium Protocols. cDNA libraries were prepared by reverse transcription, amplified with PCR, and sequenced. After alignment, matrices of gene (RNA) and protein (ADT) expression counts were processed in R 4.1.0 with Seurat, CellChat, scVelo, and FGSEA for clustering and visualization, ligand-receptor analysis, RNA velocity computation, and differential gene (DEG) expression followed by Gene Set Enrichment Analysis (GSEA).
    Figure Legend Snippet: Synthetic, experimental, and analytical overview for PUUC and MPLA+PUUC nanoparticle stimulation at early time points in the murine lung. A) Schematic of nanoparticle (NP) synthesis. PLGA NPs with or without mono-phosphoryl lipid A (MPLA) were first synthesized by double-emulsion antisolvent evaporation, followed by covalent modification with branched polyethyleneimine (PEI) with EDC/sulfo-NHS to make positively-charged PLGA-PEI NPs. Negatively charged PUUC RNA was then electrostatically loaded onto the particles to make either PUUC or MPLA+PUUC NPs. B) Schematic of experimental and analytical overview. 4 mg of PUUC and MPLA+PUUC NPs were combined with 60 μL saline and were administered into the bilateral nares of three mice per group. Doses of 20 ug PUUC and 24 ug MPLA were used per mouse. Lungs from these groups and naïve, untreated mice were harvested, processed into single-cell suspensions, and were pooled groupwise at 4 and 24 hours. Suspensions were mixed with DNA-barcoded antibodies (antibody-dependent tags; ADTs) and were processed according to 10X Chromium Protocols. cDNA libraries were prepared by reverse transcription, amplified with PCR, and sequenced. After alignment, matrices of gene (RNA) and protein (ADT) expression counts were processed in R 4.1.0 with Seurat, CellChat, scVelo, and FGSEA for clustering and visualization, ligand-receptor analysis, RNA velocity computation, and differential gene (DEG) expression followed by Gene Set Enrichment Analysis (GSEA).

    Techniques Used: Synthesized, Evaporation, Modification, Mouse Assay, Amplification, Polymerase Chain Reaction, Expressing

    7) Product Images from "Fc mediated pan-sarbecovirus protection after alphavirus vector vaccination"

    Article Title: Fc mediated pan-sarbecovirus protection after alphavirus vector vaccination

    Journal: bioRxiv

    doi: 10.1101/2022.11.28.518175

    Venezuelan Equine Encephalitis Virus Replicon Particle VRP3526 for high-titer vaccinations. A. Fragmented RNA-based assembly scheme of VRP3526 particles. B. Phylogenetic relationships of CoV spike proteins that were used in this study, including common cold CoVs (green) and prepandemic/epidemic CoVs (red). Of the β-coronaviruses, we generated spike proteins for both group 2A (HKU1) and 2B viruses. Of the group 2B viruses, we generated spike proteins for clade 1a (SARS-CoV, SHC014, WIV1), 2 (HKU3), and 1b (SARS-CoV-2, RaTG13) viruses. Tree generated from an amino acid multiple sequence alignment using Maximum Likelihood in Geneious Prime. C. VRP3526 titers obtained in this study. Dashed line denotes minimum titer required for vaccination at 2×10 4 VRP in a 10 μl footpad inoculation. D. Immunofluorescent staining at 40x magnification for VEE non-structural proteins (top) and SARS-CoV-2 spike S2 domain (middle) in Vero E6 cells infected with VRPs expressing the spike proteins used in this study.
    Figure Legend Snippet: Venezuelan Equine Encephalitis Virus Replicon Particle VRP3526 for high-titer vaccinations. A. Fragmented RNA-based assembly scheme of VRP3526 particles. B. Phylogenetic relationships of CoV spike proteins that were used in this study, including common cold CoVs (green) and prepandemic/epidemic CoVs (red). Of the β-coronaviruses, we generated spike proteins for both group 2A (HKU1) and 2B viruses. Of the group 2B viruses, we generated spike proteins for clade 1a (SARS-CoV, SHC014, WIV1), 2 (HKU3), and 1b (SARS-CoV-2, RaTG13) viruses. Tree generated from an amino acid multiple sequence alignment using Maximum Likelihood in Geneious Prime. C. VRP3526 titers obtained in this study. Dashed line denotes minimum titer required for vaccination at 2×10 4 VRP in a 10 μl footpad inoculation. D. Immunofluorescent staining at 40x magnification for VEE non-structural proteins (top) and SARS-CoV-2 spike S2 domain (middle) in Vero E6 cells infected with VRPs expressing the spike proteins used in this study.

    Techniques Used: Generated, Sequencing, Staining, Infection, Expressing

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 89
    Thermo Fisher rna
    Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rna/product/Thermo Fisher
    Average 89 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rna - by Bioz Stars, 2022-12
    89/100 stars
      Buy from Supplier

    88
    Thermo Fisher purelink rna mini kit
    Purelink Rna Mini Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/purelink rna mini kit/product/Thermo Fisher
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    purelink rna mini kit - by Bioz Stars, 2022-12
    88/100 stars
      Buy from Supplier

    99
    Thermo Fisher trizol total rna kit
    Trizol Total Rna Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trizol total rna kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trizol total rna kit - by Bioz Stars, 2022-12
    99/100 stars
      Buy from Supplier

    Image Search Results