transfection kits includes cytofect neuron transfection kit  (Cell Applications Inc)

Journal: ACS Nano

Article Title: Improved CaP Nanoparticles for Nucleic Acid and Protein Delivery to Neural Primary Cultures and Stem Cells

doi: 10.1021/acsnano.3c09608

Figure Lengend Snippet: Calcium concentration and the types of medium determine the size and quantity of CaP nanoparticles. (A–C) Different CaCl 2 concentrations caused various sizes and numbers of CaP particles. 35 mM CaCl 2 shows a great number of CaP nanoparticles with relative smaller sizes. (D,E) The example images show the transfection efficiency of pLL3.7/ Syn-Gfp under indicated CaCl 2 concentration. GFP-positive neurons were in green. The quantitative data represent the numbers of GFP-positive cells under indicated CaCl 2 concentration. (F–H) The size and quantity of CaP particles were changed by different media. Neurobasal (NB) medium has the most optimized condition to generate CaP nanoparticles. (I,J) The example images display the transfection efficiency of pLL3.7/ Syn-Gfp under various media. GFP-positive neurons were in green. The quantitative data display the numbers of GFP-positive cells under indicated CaCl 2 concentration. (K–M) The size and number of CaP nanoparticles were changed under different buffer systems. HBSS showed the most optimized condition to remove CaP nanoparticles. Over 1000 particles were analyzed from six different areas of three independent experiments. Statistic: one-way ANOVA (B, C, E, G, H, J, L, and M). Values represent the mean ± s.e.m. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The commercial transfection kits includes Cytofect Neuron Transfection Kit (Cell Applications, TF886 KS), DreamFect Transfection Reagent (OZBiosciences, DF40500), GenJet In Vitro DNA Transfection Reagent (SignaGen Laboratories, SL100488), GenMute Transfection Reagent (SignaGen Laboratories, SL100568), Lipofectamine 3000 Transfection Reagent (Thermo Fisher, L3000001), Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher, 13778075), Nupherin Transfection Reagent (Zneo, BML-SE225-0075), PolyJet Transfection Reagent (SignaGen Laboratories, SL100688), TransIT-X2 Dynamic Delivery System (Mirus Bio, MIR6003), TransIT-LT1 Transfection Reagent (Mirus Bio, MIR2304), and TurboFect Transfection Reagent (Thermo Fisher, R0531).

Techniques: Concentration Assay, Transfection

Journal: ACS Nano

Article Title: Improved CaP Nanoparticles for Nucleic Acid and Protein Delivery to Neural Primary Cultures and Stem Cells

doi: 10.1021/acsnano.3c09608

Figure Lengend Snippet: Teleofection delivers nucleic acid into primary neurons at different days in vitro (DIV) with desired efficiency. (A,B) The examples of images show the transfection efficiency of primary neurons under the indicated incubation times and CaP nanoparticle numbers. The transfected neurons were with GFP expression in green and neuronal marker MAP2 in magenta. (C,D) The quantitative data from A and B show the transfection efficiency of neurons under different transfection conditions. The transfection efficiency can be manipulated by change incubation time and loading volume of the teleofection mixture. (E) The representative images show the transfected neuron (GFP in green), glia cell marker (GFAP in red), and neuronal marker (MAP2 in magenta) at indicated time points in rat primary neuron culture. (F,G) The quantitative data of E show the transfection efficiency of neuron and glia at different DIV primary culture. (H) The quantitative data indicate the transfection efficiency of spiny and aspiny neurons under time-course manner. Data were analyzed from six different areas of three independent experiments. Statistic: one-way ANOVA (B and D) and two-way ANOVA (F and H). Values represent the mean ± s.e.m., * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The commercial transfection kits includes Cytofect Neuron Transfection Kit (Cell Applications, TF886 KS), DreamFect Transfection Reagent (OZBiosciences, DF40500), GenJet In Vitro DNA Transfection Reagent (SignaGen Laboratories, SL100488), GenMute Transfection Reagent (SignaGen Laboratories, SL100568), Lipofectamine 3000 Transfection Reagent (Thermo Fisher, L3000001), Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher, 13778075), Nupherin Transfection Reagent (Zneo, BML-SE225-0075), PolyJet Transfection Reagent (SignaGen Laboratories, SL100688), TransIT-X2 Dynamic Delivery System (Mirus Bio, MIR6003), TransIT-LT1 Transfection Reagent (Mirus Bio, MIR2304), and TurboFect Transfection Reagent (Thermo Fisher, R0531).

Techniques: In Vitro, Transfection, Incubation, Expressing, Marker

Journal: ACS Nano

Article Title: Improved CaP Nanoparticles for Nucleic Acid and Protein Delivery to Neural Primary Cultures and Stem Cells

doi: 10.1021/acsnano.3c09608

Figure Lengend Snippet: Teleofection enables versatile serial or cotransfection in primary neurons for diverse cellular biology research aims. (A,B) The flowchart shows the experiment design for serial transfection of indicated plasmids. The examples of images show the expression pattern of transfected neurons with GFP (green) or mCherry (red), and neurons were outlined by MAP2 (magenta). Under serial transfection, different nucleic acids can be uptaken by different neurons sequentially. The representative images show the neurons transfected with GFP followed by mCherry transfection at DIV10 and 11, respectively. The arrows indicate GFP and mCherry double positive neurons. (C) The quantitative data display that most of the neurons express only a single transfected gene, and only around 10% of neurons can uptake both transfected genes. (D) The representative images indicate that, based on serial-transfection stratagem, the synaptic region composed by two different neurons in green (dendritic spine) and red (axonal terminal) can be illustrated clearly. The thickness of a slice interval of a Z -stack is 0.5 μm. Also see Videos S9–S11 . (E) The flowchart shows the experiment design for serial transfection of Cmtr1 siRNA followed by pLL3.7/ Syn-Gfp transfection at DIV7 and 10, respectively. The neurons were fixed for ICC at DIV20. (F) The representative images show the neuron-expressed GFP (green) and mCherry (red) as the control and Cmtr1 knockdown group, respectively. The CMTR1 signal was displayed as magenta. (G) The high-magnification images from F show the details of transfected neurons. (H–J) The quantitative data from F and G represent CMTR1 intensity, dendritic number, and dendritic length in control and Cmtr1 siRNA knockdown groups. (K) The cartoon displays the definition and the pattern diagram of the dendritic branch types for image analysis. The quantitative data display the percentage of dendritic branch types in control and Cmtr1 siRNA-transfected neurons. Data were analyzed from at least six different areas of three independent experiments. Statistic: Student’s unpaired t test (H–K). Values represent the mean ± s.e.m., * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The commercial transfection kits includes Cytofect Neuron Transfection Kit (Cell Applications, TF886 KS), DreamFect Transfection Reagent (OZBiosciences, DF40500), GenJet In Vitro DNA Transfection Reagent (SignaGen Laboratories, SL100488), GenMute Transfection Reagent (SignaGen Laboratories, SL100568), Lipofectamine 3000 Transfection Reagent (Thermo Fisher, L3000001), Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher, 13778075), Nupherin Transfection Reagent (Zneo, BML-SE225-0075), PolyJet Transfection Reagent (SignaGen Laboratories, SL100688), TransIT-X2 Dynamic Delivery System (Mirus Bio, MIR6003), TransIT-LT1 Transfection Reagent (Mirus Bio, MIR2304), and TurboFect Transfection Reagent (Thermo Fisher, R0531).

Techniques: Cotransfection, Transfection, Expressing, Control, Knockdown

Journal: ACS Nano

Article Title: Improved CaP Nanoparticles for Nucleic Acid and Protein Delivery to Neural Primary Cultures and Stem Cells

doi: 10.1021/acsnano.3c09608

Figure Lengend Snippet: Teleofection is applicable for molecular and biochemical analysis. (A) The flowchart shows the experiment design for cotransfection of pLL3.7/ Syn-Gfp and -mCherry at DIV7. Transfected neurons were harvested at DIV10 for Western blotting to detect the expression of GFP and mCherry. (B) The representative images show the protein expression level of delivery genes ( Gfp and mCherry ) under dose- and time-dependent manners in primary neurons. Two independent samples were displayed for each condition. Tubulin, actin, and GAPDH were detected as internal controls. (C) The flowchart shows the experiment design for Gfp mRNA transfection at DIV10, followed by sample collection at indicated times. (D) The representative image shows the expression level of GFP under time-dependent manners in primary neurons. (E) The flowchart indicates the experiment design for a dual-reporter assay in primary neurons. Firefly and Renilla Luciferase reporter plasmids were cotransfected into neurons at DIV7, and the reporter assay was performed at DIV10. (F) The expression of endogenous CPEB2 was detected by Western blotting. (G,H) The table represents the Luciferase activity of Firefly and Renilla in transfected neurons. The quantitative data were displayed as a dot plot graph in H. Data were analyzed from three independent experiments. Statistic: Student’s unpaired t test (H). Data were analyzed from three independent experiments. Values represent the mean ± s.e.m., * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The commercial transfection kits includes Cytofect Neuron Transfection Kit (Cell Applications, TF886 KS), DreamFect Transfection Reagent (OZBiosciences, DF40500), GenJet In Vitro DNA Transfection Reagent (SignaGen Laboratories, SL100488), GenMute Transfection Reagent (SignaGen Laboratories, SL100568), Lipofectamine 3000 Transfection Reagent (Thermo Fisher, L3000001), Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher, 13778075), Nupherin Transfection Reagent (Zneo, BML-SE225-0075), PolyJet Transfection Reagent (SignaGen Laboratories, SL100688), TransIT-X2 Dynamic Delivery System (Mirus Bio, MIR6003), TransIT-LT1 Transfection Reagent (Mirus Bio, MIR2304), and TurboFect Transfection Reagent (Thermo Fisher, R0531).

Techniques: Cotransfection, Transfection, Western Blot, Expressing, Reporter Assay, Luciferase, Activity Assay

Journal: ACS Nano

Article Title: Improved CaP Nanoparticles for Nucleic Acid and Protein Delivery to Neural Primary Cultures and Stem Cells

doi: 10.1021/acsnano.3c09608

Figure Lengend Snippet: Teleofection is a stratagem for protein delivery in cell lines and primary neuron culture. (A) The flowchart shows the experiment design to generate different CaP nanoparticles with distinct proteins for the cell delivery. Also see Supporting Information Figure S8 for the stratagem of protein cotransfection by teleofection. (B) The representative images showcase nanoparticles containing various fluorescent dye-conjugated antimouse antibodies (Ab), showing green for Ab-488, red for Ab-594, and magenta for Ab-647. (C) The representative images show the global amount of CTTNBP2-GFP (green) in COS7 cells after protein transfection by teleofection. The high magnification displays the changes of CTTNBP2-GFP in a COS7 cell under time-dependent manner. The bright field displays the details of cell morphology. (D–E) The dot plot graphs represent the quantitative data of transfection efficiency and the degradation rate of CTTNBP2-GFP in COS7 cells. (F) The example of images indicates the distribution of CTTNBP2-GFP (green) in COS7 cells under different cell cycle stages. Nuclei and stress fibers were outlined by DAPI (blue) and phalloidin (red), respectively. The colocalization of CTTNBP2-GFP and stress fiber was displayed as yellow. (G) The flowchart indicates the experiment design to deliver reporter gene and protein sequentially in primary neuron culture. (H) The representative images show the distribution of CTTNBP2-GFP (green) in primary neurons after 24 h of transfection. The low-power field displays the global view of the primary neuron labeled by mCherry (red). Nuclei were labeled by DAPI in blue. The high magnification reveals the detailed distribution of CTTNBP2-GFP in the dendritic shaft. The colocalization of CTTNBP2-GFP and mCherry was manifested as yellow. (I) The representative images show the distribution of NuMA-C-GFP (green) in primary neurons after 24 h of teleofection. The low-power field displays the global view of primary neurons labeled by mCherry (white); nuclei were displayed as red by DAPI. The high magnification shows the distribution of NuMA-C-GFP in nuclei; the colocalization of NuMA-C-GFP and the nucleus was manifested as yellow. Data were analyzed from six different areas of three independent experiments. Statistic: One-way ANOVA (D and E). Values represent the mean ± s.e.m., * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The commercial transfection kits includes Cytofect Neuron Transfection Kit (Cell Applications, TF886 KS), DreamFect Transfection Reagent (OZBiosciences, DF40500), GenJet In Vitro DNA Transfection Reagent (SignaGen Laboratories, SL100488), GenMute Transfection Reagent (SignaGen Laboratories, SL100568), Lipofectamine 3000 Transfection Reagent (Thermo Fisher, L3000001), Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher, 13778075), Nupherin Transfection Reagent (Zneo, BML-SE225-0075), PolyJet Transfection Reagent (SignaGen Laboratories, SL100688), TransIT-X2 Dynamic Delivery System (Mirus Bio, MIR6003), TransIT-LT1 Transfection Reagent (Mirus Bio, MIR2304), and TurboFect Transfection Reagent (Thermo Fisher, R0531).

Techniques: Cotransfection, Transfection, Labeling

Journal: ACS Nano

Article Title: Improved CaP Nanoparticles for Nucleic Acid and Protein Delivery to Neural Primary Cultures and Stem Cells

doi: 10.1021/acsnano.3c09608

Figure Lengend Snippet: Teleofection introduces an innovative method for transporting both nucleic acids and proteins into NSCs. (A) The flowchart shows the experiment design to deliver nucleic acid into NSCs by teleofection. (B,C) The dot plot graph represents the quantitative data of transfection efficiency of pLL3.7/ Syn-Gfp in NSCs. The representative images show the population of GFP-positive cells after teleofection. The high-power field exhibits the coexpression of NSC markers, Nestin and Notch1, in GFP-positive cells. (D) The example images show the expression level of GFP under dose-dependent manners by Western blotting after teleofection in NSCs. (E) The table represents the activity of Firefly and Rellina Luciferase in transfected NSCs. The quantitative data were displayed as a dot plot graph. Data were analyzed from four independent experiments. (F) The dot plot graph displays the transfection efficiency of CTTNBP2-GFP in NSCs by teleofection with a dose-dependent manner. (G) The representative images show the distribution of CTTNBP2-GFP (green) in NSCs after 24 h of teleofection. Nuclei and F-actin were labeled by DAPI and phalloidin in blue and red, respectively. The colocalization of CTTNBP2-GFP and F-actin was manifested as yellow. The orthogonal view of white squares from NSCs provides a comprehensive understanding of the relative distribution of CTTNBP2-GFP and F-actin from multiple angles. Statistic: One-way ANOVA (B and F). Student’s unpaired t test (E). Values represent the mean ± s.e.m., * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The commercial transfection kits includes Cytofect Neuron Transfection Kit (Cell Applications, TF886 KS), DreamFect Transfection Reagent (OZBiosciences, DF40500), GenJet In Vitro DNA Transfection Reagent (SignaGen Laboratories, SL100488), GenMute Transfection Reagent (SignaGen Laboratories, SL100568), Lipofectamine 3000 Transfection Reagent (Thermo Fisher, L3000001), Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher, 13778075), Nupherin Transfection Reagent (Zneo, BML-SE225-0075), PolyJet Transfection Reagent (SignaGen Laboratories, SL100688), TransIT-X2 Dynamic Delivery System (Mirus Bio, MIR6003), TransIT-LT1 Transfection Reagent (Mirus Bio, MIR2304), and TurboFect Transfection Reagent (Thermo Fisher, R0531).

Techniques: Transfection, Expressing, Western Blot, Activity Assay, Luciferase, Labeling

Journal: Frontiers in Cell and Developmental Biology

Article Title: MicroRNA-494 Regulates Endoplasmic Reticulum Stress in Endothelial Cells

doi: 10.3389/fcell.2021.671461

Figure Lengend Snippet: ER stress induces expression of the primary and mature forms of miR-494. Relative mRNA expression of ER stress responsive genes as measured using qRT-PCR. (A) spliced XBP1 and Total XBP1 and (B) DDIT3 (CHOP) mRNA levels in HUVECs treated with 5 μg/mL tunicamycin (TCN) over a time course. Relative expression of (C) mature miR-494 and (D) primary miR-494 (pri-miR-494) in HUVECs treated with TCN over a time course. Gene expression is normalized to GAPDH or U6 and mean fold change compared to vehicle control or time 0h is shown. Graphs are representative of one biological replicate from three independent replicates where values indicate mean ± standard deviation.

Article Snippet: For plasmid transfections, HUVECs were transfected using the Cytofect HUVECs transfection kit (Cat:TF200K, Cell Applications). pEGFP-ATF6 was a gift from Ron Prywes (Addgene plasmid # 32955) ( ).

Techniques: Expressing, Quantitative RT-PCR, Gene Expression, Control, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: MicroRNA-494 Regulates Endoplasmic Reticulum Stress in Endothelial Cells

doi: 10.3389/fcell.2021.671461

Figure Lengend Snippet: miR-494 is a negative regulator of ER stress in vitro . Relative mRNA expression of ER stress responsive genes as measured by qRT-PCR. (A,B) DDIT3 (CHOP), (C,D) spliced XBP1 in HUVECs treated with 10 μg/mL TCN 48h after transfection with (A,C) miR-494 mimic or (B,D) miR-494 inhibitor. Gene expression is normalized to GAPDH and mean fold changes compared to control treatments are shown. (E,F) Simple Western blot analysis of HUVECs transfected with miR-494 mimic or control (24 h) (E) or miR-494 inhibitor or control (F) followed by TCN (10 μg/mL) for 24 h. (G) Cell viability in HUVECs as treated in (A) Vertical dotted red line indicates non-adjacent lanes. Graphs are mean + SEM fold changes of biological replicates from n = 3 independent experiments. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 by two-tailed Student’s T -test.

Article Snippet: For plasmid transfections, HUVECs were transfected using the Cytofect HUVECs transfection kit (Cat:TF200K, Cell Applications). pEGFP-ATF6 was a gift from Ron Prywes (Addgene plasmid # 32955) ( ).

Techniques: In Vitro, Expressing, Quantitative RT-PCR, Transfection, Gene Expression, Control, Simple Western, Two Tailed Test

Journal: Frontiers in Cell and Developmental Biology

Article Title: MicroRNA-494 Regulates Endoplasmic Reticulum Stress in Endothelial Cells

doi: 10.3389/fcell.2021.671461

Figure Lengend Snippet: miR-494 regulates target genes in cell survival and DNA replication. (A) Venn diagram showing the number of downregulated target proteins in a Tandem Mass Tag labeled Mass Spectrometry profile from HUVECs treated with TCN or transfected with miR-494 compared to vehicle treatment or control miR respectively. (B) Fold-change (compared to respective controls) of protein or mRNA levels as assessed by Mass Spectrometry or qRT-PCR respectively for the six targets that were downregulated in both TCN and miR-494 groups. All six targets harbor miR-494 binding sites in their 3′UTRs. (C) Representative Simple Western blot showing survivin ( BIRC5 ) and GINS4 levels in HUVECs 24 h after miR-494 transfection followed by TCN treatment (24 h). Right panels show quantitation of biological replicates. ∗ P < 0.05, ∗∗ P < 0.01, by two-tailed Student’s T -test. (D) Immunofluorescence images showing survivin expression in HUVECs 24 h after miR transfection and/or TCN treatment. Right panel shows quantification via ImageJ of survivin fluorescence intensity in each cell. Each dot represents individual cells. Scale bar in white = 50 μm. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 by one-way ANOVA with post hoc Tukey’s correction.

Article Snippet: For plasmid transfections, HUVECs were transfected using the Cytofect HUVECs transfection kit (Cat:TF200K, Cell Applications). pEGFP-ATF6 was a gift from Ron Prywes (Addgene plasmid # 32955) ( ).

Techniques: Labeling, Mass Spectrometry, Transfection, Control, Quantitative RT-PCR, Binding Assay, Simple Western, Quantitation Assay, Two Tailed Test, Immunofluorescence, Expressing, Fluorescence

Journal: Nature communications

Article Title: Stress-induced RNA-chromatin interactions promote endothelial dysfunction.

doi: 10.1038/s41467-020-18957-w

Figure Lengend Snippet: Fig. 1 High glucose and TNFα induce a profound gene expression and phenotypic change. a HUVECs were treated in biological duplicates as Day 0: 25 mM mannitol as normal glucose and osmolarity control (NM); Day 3: combined treatment consisting 25 mM D-glucose and 5 ng/mL TNFα (H + T) for 3 days; and Day 7: H + T treatment for 7 days. Each group of treated cells was subjected to single-cell RNA-seq (scRNA-seq), Hi-C, and iMARGI assays. b t-SNE plot of scRNA-seq (4000–15,000 cells per sample) showed clear separation by treatment condition into three distinct clusters. c Principal component analysis of scRNA-seq data: single cells are plotted in the first two PC space and are labeled in red (Day 0, i.e., NM), green (Day 3, i.e., 3-day H + T treatment), and blue (Day 7, i.e., 7-day H + T treatment). d Expression heatmap (z-scaled) of top DE genes in single ECs grouped into functional pathways. Cells were ordered by increasing SERPINE1 expression (per each sample separately) and binned per 100 cells for the analysis. A total of 269 bins in Day 0, 177 bins in Day 3, and 148 bins in Day 7. e t-SNE plots of the expression level of selected genes in each single cell across the time course. The RNA levels are represented by log-normalized unique molecular identifier counts. f mRNA levels of eNOS and α-SMA in NM vs. H + T-treated HUVECs and cells untreated (NT) or treated with TGF-β (10 ng/mL) and IL-1β (5 ng/mL; T + I) for 3 or 7 days. The respective control was set as 1. Relative eNOS level: data represent mean ± SEM from five independent experiments; relative α-SMA level in H + T treatment: data represent mean ± SEM from seven independent experiments; relative α-SMA level in T + I treatment: data represent mean ± SEM from four independent experiments. *P = 0.0067, 0.0087, 0.0057, and 0.0017 from left to right based on ANOVA with Bonferroni as post hoc test. g Cell morphology under bright field (BF), immunofluorescent staining of α-SMA, and VE-cadherin (VE-cad), phalloidin staining of cytoskeleton, and DRAQ5 (DRAQ) staining of the nuclei. Representative images from five independent experiments are shown. Scale bar of BF = 100 µm; scale bars of (immuno)fluorescent staining = 50 µm. Source data are provided as a Source data file.

Article Snippet: Plasmid transfection was performed using the CytofectTM HUVEC Transfection kit (Cell Applications) following the manufacturer’s protocol in 6-well or 12-well plates.

Techniques: Gene Expression, Control, RNA Sequencing, Hi-C, Labeling, Expressing, Functional Assay, Staining

Journal: Nature communications

Article Title: Stress-induced RNA-chromatin interactions promote endothelial dysfunction.

doi: 10.1038/s41467-020-18957-w

Figure Lengend Snippet: Fig. 2 Overview of time-course Hi-C and iMARGI data. a, b Proportions of intrachromosomal (yellow) and interchromosomal read pairs (blue) in Hi-C (a) and iMARGI data (b) at the three time points (columns). c An example of interchromosomal iMARGI read pairs mapped to chromosome 2 near LINC00607 (left) and chromosome 7 near SERPINE1 (right). The RNA end (pink) and the DNA end (green) of each read pair is linked by a horizontal line. d Examples of overlapping iMARGI read pairs on a contact matrix from the RNA end (rows) to the DNA end (columns) with SEs (marked in light blue and in SE tracks) in control (Day 0) ECs. Genome region: chr1:75,000,0000–chr1:125,000,000. Resolution = 200 kb. e Proportions of iMARGI read pairs with the RNA ends (pink) or the DNA ends (green) in Day 0 (Ctrl) and Days 3 and 7 (H + T) ECs mapped to HUVEC SEs. Dotted line: the relative size of HUVEC SEs compared to the size of the genome. Source data are provided as a Source data file.

Article Snippet: Plasmid transfection was performed using the CytofectTM HUVEC Transfection kit (Cell Applications) following the manufacturer’s protocol in 6-well or 12-well plates.

Techniques: Hi-C, Control

Journal: Nature communications

Article Title: Stress-induced RNA-chromatin interactions promote endothelial dysfunction.

doi: 10.1038/s41467-020-18957-w

Figure Lengend Snippet: Fig. 4 Inhibition of LINC00607 attenuates SERPINE1 induction and EC dysfunction. a Illustration of LINC00607 genomic locus and gene structure and LNA GapmeRs targeting of LINC00607 RNA. b HUVECs were transfected with scramble (scr) or two LNAs targeting LINC00607. LINC00607 RNA and SERPINE1 mRNA levels were quantified. Data represent mean ± SEM from seven independent experiments. c qPCR of LINC00607 in subcellular fractionations of HUVECs transfected with scr, LNA1, or LNA2. Data represent mean ± SEM from four independent experiments. d RNA-seq was performed with cells transfected as in b in biological replicates. Heatmap is plotted based on z-scaled log-transformed gene expression levels. e, f ECs transfected with scr or LNA1 and then treated by H + T were used in e monocyte adhesion assay and in f SA-β-gal assay. In e, the number of attached monocytes to ECs were quantified. Data represent mean ± SEM from eight independent experiments performed with peripheral blood-derived monocytes from four individual donors. Representative images show the attachment of fluorescently labeled peripheral blood-derived monocytes to ECs for experiments performed with monocytes from three different donors. In f, ECs with positive β-gal staining were quantified. The positively stained cell number in scr control was set to 100 (%). f Data represent mean ± SEM from four independent experiments respectively. Scale bar = 200 μm. * denotes p = 0.0289, 0.0013, 0.001, 0.0001, 0.0025, and 0.024 from left to right (in b) and p = 0.0023, 0.0079, and 0.0249 from left to right (in c) between indicated groups based on ANOVA followed by Bonferroni post hoc test (in b and c); p = 0.0039 (in e) and 0.0014 (in f), as compared to scr group based on two-tailed paired t test (in e and f). Source data are provided as a Source data file.

Article Snippet: Plasmid transfection was performed using the CytofectTM HUVEC Transfection kit (Cell Applications) following the manufacturer’s protocol in 6-well or 12-well plates.

Techniques: Inhibition, Transfection, RNA Sequencing, Transformation Assay, Gene Expression, Cell Adhesion Assay, Derivative Assay, Labeling, Staining, Control, Two Tailed Test