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  • 99
    Integrated DNA Technologies electroporation enhancer
    Electroporation Enhancer, supplied by Integrated DNA Technologies, used in various techniques. Bioz Stars score: 99/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher electroporation cuvette
    miR-195 and miR-497 decrease osteogenesis in human primary MSC A. miR-195 and miR-497 levels after <t>electroporation</t> of human MSC with miR-195 and miR-497 were determined by quantitative real time PCR. Small nuclear RNA U6 was used as reference gene. Values shown represent 2 independent experiments and are relative to scrambled negative control (SCR). B. ALP staining 7 days after electroporation of human MSC with scrambled negative control (SCR), miR-195 or miR-497 (5X; microscope scale: 50 μm; photography scale: 2 mm). C. ALP and RUNX2 expression levels in human MSC electroporated with either SCR, miR-195, miR-497. D. ALP expression levels 48h after MSC transfection with SCR negative control or anti-miR-195; ALP staining quantification in MSC transfected with SCR negative control or anti-miR-195 after 5 days of incubation with osteogenic supplements. Values shown represent 2 independent experiments and are relative to SCR (mean±SD; * P
    Electroporation Cuvette, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 140 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher electroporation
    miR-195 and miR-497 decrease osteogenesis in human primary MSC A. miR-195 and miR-497 levels after <t>electroporation</t> of human MSC with miR-195 and miR-497 were determined by quantitative real time PCR. Small nuclear RNA U6 was used as reference gene. Values shown represent 2 independent experiments and are relative to scrambled negative control (SCR). B. ALP staining 7 days after electroporation of human MSC with scrambled negative control (SCR), miR-195 or miR-497 (5X; microscope scale: 50 μm; photography scale: 2 mm). C. ALP and RUNX2 expression levels in human MSC electroporated with either SCR, miR-195, miR-497. D. ALP expression levels 48h after MSC transfection with SCR negative control or anti-miR-195; ALP staining quantification in MSC transfected with SCR negative control or anti-miR-195 after 5 days of incubation with osteogenic supplements. Values shown represent 2 independent experiments and are relative to SCR (mean±SD; * P
    Electroporation, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3150 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad electroporation
    miR-195 and miR-497 decrease osteogenesis in human primary MSC A. miR-195 and miR-497 levels after <t>electroporation</t> of human MSC with miR-195 and miR-497 were determined by quantitative real time PCR. Small nuclear RNA U6 was used as reference gene. Values shown represent 2 independent experiments and are relative to scrambled negative control (SCR). B. ALP staining 7 days after electroporation of human MSC with scrambled negative control (SCR), miR-195 or miR-497 (5X; microscope scale: 50 μm; photography scale: 2 mm). C. ALP and RUNX2 expression levels in human MSC electroporated with either SCR, miR-195, miR-497. D. ALP expression levels 48h after MSC transfection with SCR negative control or anti-miR-195; ALP staining quantification in MSC transfected with SCR negative control or anti-miR-195 after 5 days of incubation with osteogenic supplements. Values shown represent 2 independent experiments and are relative to SCR (mean±SD; * P
    Electroporation, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 11432 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad electroporation a micropulser electroporator
    miR-195 and miR-497 decrease osteogenesis in human primary MSC A. miR-195 and miR-497 levels after <t>electroporation</t> of human MSC with miR-195 and miR-497 were determined by quantitative real time PCR. Small nuclear RNA U6 was used as reference gene. Values shown represent 2 independent experiments and are relative to scrambled negative control (SCR). B. ALP staining 7 days after electroporation of human MSC with scrambled negative control (SCR), miR-195 or miR-497 (5X; microscope scale: 50 μm; photography scale: 2 mm). C. ALP and RUNX2 expression levels in human MSC electroporated with either SCR, miR-195, miR-497. D. ALP expression levels 48h after MSC transfection with SCR negative control or anti-miR-195; ALP staining quantification in MSC transfected with SCR negative control or anti-miR-195 after 5 days of incubation with osteogenic supplements. Values shown represent 2 independent experiments and are relative to SCR (mean±SD; * P
    Electroporation A Micropulser Electroporator, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore electroporation
    miR-195 and miR-497 decrease osteogenesis in human primary MSC A. miR-195 and miR-497 levels after <t>electroporation</t> of human MSC with miR-195 and miR-497 were determined by quantitative real time PCR. Small nuclear RNA U6 was used as reference gene. Values shown represent 2 independent experiments and are relative to scrambled negative control (SCR). B. ALP staining 7 days after electroporation of human MSC with scrambled negative control (SCR), miR-195 or miR-497 (5X; microscope scale: 50 μm; photography scale: 2 mm). C. ALP and RUNX2 expression levels in human MSC electroporated with either SCR, miR-195, miR-497. D. ALP expression levels 48h after MSC transfection with SCR negative control or anti-miR-195; ALP staining quantification in MSC transfected with SCR negative control or anti-miR-195 after 5 days of incubation with osteogenic supplements. Values shown represent 2 independent experiments and are relative to SCR (mean±SD; * P
    Electroporation, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 504 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Amaxa conventional electroporation
    Efficiency of <t>electroporation</t> and cell recovery using different methods. Naïve T-cells electroporated with an mRNA for the fluorescent protein Ruby. ( a ) Percentage of Ruby positive cells following electroporation using three different commercially available electroporators: BTX, <t>Amaxa</t> from Lonza and Neon from ThermoFisher. U-14, V-23 and T-23 are electroporation setting on the Amaxa machine. 1700 and 2150 are the voltages used on the Neon (see Materials and methods for more details). ( b ) Number of cell recovered following electroporation- a product of cell viability and cell count compared to starting condition (dashed grey line).
    Conventional Electroporation, supplied by Amaxa, used in various techniques. Bioz Stars score: 86/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad electroporation buffer
    Molecular ( a – c ) and biological ( d – f ) changes induced by Echinococcus granulosus Calmodulin (EgCaM)-specific siRNA using three delivery methods in different developmental stages in vitro . ( a – c ) EgCaM expression profile in different in vitro stages: protoscoleces (PSC) 3 and 8 days after <t>electroporation</t> (EP), soaking (SK) and electro-soaking (ES); microcysts (MC) and strobilated worms (SW). ( d ) Viability changes of protoscoleces treated with EgCaM-specific siRNA. ( e ) Size changes of 30 protoscoleces treated with EgCaM-specific siRNA. ( f ) Changes in body contractions per minute in the strobilated worms treated with EgCaM-specific siRNA. Data was compared to the controls: control protoscoleces (Ctrl) and negative siRNA control (siR-Ctrl). The gene expression data for siR-Ctrl were only demonstrated for electro-soaking method. Bars show the mean ± standard deviation (SD) derived from duplicates experiments. (**P
    Electroporation Buffer, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 486 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Lonza electroporation buffer
    Molecular ( a – c ) and biological ( d – f ) changes induced by Echinococcus granulosus Calmodulin (EgCaM)-specific siRNA using three delivery methods in different developmental stages in vitro . ( a – c ) EgCaM expression profile in different in vitro stages: protoscoleces (PSC) 3 and 8 days after <t>electroporation</t> (EP), soaking (SK) and electro-soaking (ES); microcysts (MC) and strobilated worms (SW). ( d ) Viability changes of protoscoleces treated with EgCaM-specific siRNA. ( e ) Size changes of 30 protoscoleces treated with EgCaM-specific siRNA. ( f ) Changes in body contractions per minute in the strobilated worms treated with EgCaM-specific siRNA. Data was compared to the controls: control protoscoleces (Ctrl) and negative siRNA control (siR-Ctrl). The gene expression data for siR-Ctrl were only demonstrated for electro-soaking method. Bars show the mean ± standard deviation (SD) derived from duplicates experiments. (**P
    Electroporation Buffer, supplied by Lonza, used in various techniques. Bioz Stars score: 99/100, based on 83 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher electroporation buffer
    GFP and VEGF gene expression of electroporated BMMSCs. Green fluorescence represents the successful <t>electroporation</t> of plasmid into BMMSCs. Electroporated cells after 48 h ( a and b ). G418 selection after 20 days ( d and e ) ( bright vision
    Electroporation Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 251 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Harvard Bioscience electroporation chamber
    Robo/Dll1 Signaling Regulates the Balance between Direct Neurogenesis and IPC Abundance in Chick Dorsal Pallium (A–D) Analysis of chick dorsal pallium at 6 days post-ovoposition (dpo), showing many neurons in the VZ (solid arrowheads) in the medial part (mDP; B) and basal mitoses (open arrowheads) in the lateral (lDP; C; n = 3 embryos; t tests). (E–H) ISH in chick DP at 6 dpo, and quantifications of intensity (au, arbitrary units). High magnifications show chkRobo1 (E’ and E”) and chkDll1 (G’ and G”) in the indicated regions. Panel shown in (G) is a tiled image. (I) qPCR analysis in the VZ of the regions and species indicated. Values are ratio Robo1 to Gapdh (n = 12–15 replicates; paired or independent samples t tests). (J–O) Experimental design to manipulate in ovo Robo and Dll1 in mDP and lDP, representative examples and quantifications of neurons in the VZ (Tbr1+) and basal mitoses (PH3, open arrowheads; n = 3-5 embryos per group; t tests in L and Q; one-way ANOVA followed by t tests in M and O). Red shadowing and asterisks in (L) and (O) indicate values within group corresponding to GFP− cells. (P and Q) Expression of Tbr2 in basal PH3+ mitoses (solid arrowhead, Tbr2+; open arrowhead, Tbr2−) upon <t>electroporation</t> of dnR1/2+Dll1 in mDP (as in J and K), and quantification (n = 4–5 embryos; t tests). (R–U) Analysis of neuronal clones (GFP+RFP+Tuj1+) upon overexpression of dnR1/2+Dll1+Gfp, representative examples and quantification (n = 52 clones Gfp, 59 clones dnR1/2+Dll1+Gfp, 3–7 embryos; t test or χ2-test). Values are mean + SEM; ∗ p
    Electroporation Chamber, supplied by Harvard Bioscience, used in various techniques. Bioz Stars score: 91/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad electroporation cuvette
    Electric field enhancement by CNT in <t>electroporation.</t> (a) Sketch of electroporation with CNT-free (left) and CNT associated cells (right). Table insert: theoretical electric field ( E ) enhancement over the applied field ( E 0 ) by a nanotube of 1 μ m long. d denotes the distance to nanotube tip. a 1 and a 2 are the aspect ratios (i.e. diameter/length) of nanotubes, and equal to 0.1 and 0.5 respectively. (b) Electroporation time constants versus the voltages. A series of time constants of the CNT associated Bal17 cells were obtained under various voltages. They were compared to the time constant of regular Bal17 cells at 230 V. (c) Improvement of EGFP transfection by CNT mediated electroporation in Bal17 cells. The gates of EGFP positive cells were adjusted based on the Neg Contl and Pos Contl . The mean values of EGFP levels (left axis) and the percentages (right axis) of gated viable cells are shown. Neg Contl : negative control, cells speared and electroporated without plasmid; Pos Contl : positive control, cells without spearing and electroporated at 230 V with EGFP plasmid; 180, 200 and 220 V: CNT associated cells and electroporated with EGFP plasmids at corresponding voltages.
    Electroporation Cuvette, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 1604 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Eppendorf AG electroporation cuvette
    CRF protein homo- and heterodimerization. (A) CRF proteins CRF1–CRF8 and just the CRF domain of CRF2 (otherwise noted as CRFD) were analysed for potential homo- and heterodimerization using both BiFC and Y2H. Positive interactions are noted as a (+), (+*) for both BiFC and Y2H together, and (–) for non-interactions. (B) A cartoon of the conserved regions of CRF1–CRF6 proteins, the highly related CRF7 and CRF8 proteins (lacking the C-terminal third of the protein, but containing both the CRF domain and the AP2/ERF domain), and CRFD (just the CRF domain of CRF2). (C, D) Representative examples of positive CRF protein homo- and heterodimerization by BiFC as indicated in A are shown both under UV light in the presence of Hoechst 33342 dye denoting the nucleus and using a YFP wavelength filter to visualize BiFC interaction. Additionally, representative examples of empty vector (EV) controls for both N- and C-terminal BiFC vectors (EVn and EVc) examined in A are shown versus various CRFs and each other. (C) BiFC interactions in protoplasts transformed via <t>electroporation.</t> (D) BiFC interactions in tobacco leaves transformed via Agrobacterium .
    Electroporation Cuvette, supplied by Eppendorf AG, used in various techniques. Bioz Stars score: 99/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Amaxa electroporation cuvettes
    CRF protein homo- and heterodimerization. (A) CRF proteins CRF1–CRF8 and just the CRF domain of CRF2 (otherwise noted as CRFD) were analysed for potential homo- and heterodimerization using both BiFC and Y2H. Positive interactions are noted as a (+), (+*) for both BiFC and Y2H together, and (–) for non-interactions. (B) A cartoon of the conserved regions of CRF1–CRF6 proteins, the highly related CRF7 and CRF8 proteins (lacking the C-terminal third of the protein, but containing both the CRF domain and the AP2/ERF domain), and CRFD (just the CRF domain of CRF2). (C, D) Representative examples of positive CRF protein homo- and heterodimerization by BiFC as indicated in A are shown both under UV light in the presence of Hoechst 33342 dye denoting the nucleus and using a YFP wavelength filter to visualize BiFC interaction. Additionally, representative examples of empty vector (EV) controls for both N- and C-terminal BiFC vectors (EVn and EVc) examined in A are shown versus various CRFs and each other. (C) BiFC interactions in protoplasts transformed via <t>electroporation.</t> (D) BiFC interactions in tobacco leaves transformed via Agrobacterium .
    Electroporation Cuvettes, supplied by Amaxa, used in various techniques. Bioz Stars score: 91/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Avantor electroporation cuvettes
    Cerebellar organoids <t>electroporation</t> with Otx2 / c-MYC induces Group 3 MB in vivo. a Schematic representation of in vivo injection of modified cerebellar organoids. b DAPI staining and GFP immunofluorescence (Venus) of the sagittal brain section of nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. c Confocal images of GFP (Venus) and PCNA immunofluorescence of tumors in nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. d NPR3 immunohistochemistry and Hematoxylin staining of tumor in nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. e MDS (multidimensional scaling) analysis performed on the 1000 most variable probes of the whole-genome DNA methylation data shows a close similarity between organoids and group 3 MBs. Color legend of the MDS plot as follows: OM Organoids (Organoids_OM, black); GM Organoids (Organoids_GM, gray); WNT, Wingless MB (blue); SHH-A, Sonic Hedgehog MB-adulthood and childhood (red); SHH-B Sonic Hedgehog MB infant (dark red); G3, Group 3 MB (yellow); G4, Group 4 MB (green). f Hierarchical clustering and heatmap of beta values relative to the 39 high-quality CpG islands better discriminating MB subgroup in the Hovestadt set (Hovestadt et al.) 35 . The heatmap shows normalized methylation levels in organoid samples and MB samples. Clusters were obtained by means of Ward’s minimum variance method, using the Euclidean distance. Color legend: OM Organoids (Organoids_OM, black); GM Organoids (Organoids_ GM, gray); WNT, Wingless MB (blue); SHH-A, Sonic Hedgehog MB-adulthood and childhood (red); SHH-B Sonic Hedgehog MB infant (dark red); G3, Group 3 MB (yellow); G4, Group 4 MB (green). Scale bars 1 mm in ( b ), 100 µm in ( c , d ).
    Electroporation Cuvettes, supplied by Avantor, used in various techniques. Bioz Stars score: 94/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Kaneka Corp electroporation cuvettes
    Cerebellar organoids <t>electroporation</t> with Otx2 / c-MYC induces Group 3 MB in vivo. a Schematic representation of in vivo injection of modified cerebellar organoids. b DAPI staining and GFP immunofluorescence (Venus) of the sagittal brain section of nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. c Confocal images of GFP (Venus) and PCNA immunofluorescence of tumors in nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. d NPR3 immunohistochemistry and Hematoxylin staining of tumor in nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. e MDS (multidimensional scaling) analysis performed on the 1000 most variable probes of the whole-genome DNA methylation data shows a close similarity between organoids and group 3 MBs. Color legend of the MDS plot as follows: OM Organoids (Organoids_OM, black); GM Organoids (Organoids_GM, gray); WNT, Wingless MB (blue); SHH-A, Sonic Hedgehog MB-adulthood and childhood (red); SHH-B Sonic Hedgehog MB infant (dark red); G3, Group 3 MB (yellow); G4, Group 4 MB (green). f Hierarchical clustering and heatmap of beta values relative to the 39 high-quality CpG islands better discriminating MB subgroup in the Hovestadt set (Hovestadt et al.) 35 . The heatmap shows normalized methylation levels in organoid samples and MB samples. Clusters were obtained by means of Ward’s minimum variance method, using the Euclidean distance. Color legend: OM Organoids (Organoids_OM, black); GM Organoids (Organoids_ GM, gray); WNT, Wingless MB (blue); SHH-A, Sonic Hedgehog MB-adulthood and childhood (red); SHH-B Sonic Hedgehog MB infant (dark red); G3, Group 3 MB (yellow); G4, Group 4 MB (green). Scale bars 1 mm in ( b ), 100 µm in ( c , d ).
    Electroporation Cuvettes, supplied by Kaneka Corp, used in various techniques. Bioz Stars score: 90/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Amaxa electroporation machine
    Cerebellar organoids <t>electroporation</t> with Otx2 / c-MYC induces Group 3 MB in vivo. a Schematic representation of in vivo injection of modified cerebellar organoids. b DAPI staining and GFP immunofluorescence (Venus) of the sagittal brain section of nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. c Confocal images of GFP (Venus) and PCNA immunofluorescence of tumors in nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. d NPR3 immunohistochemistry and Hematoxylin staining of tumor in nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. e MDS (multidimensional scaling) analysis performed on the 1000 most variable probes of the whole-genome DNA methylation data shows a close similarity between organoids and group 3 MBs. Color legend of the MDS plot as follows: OM Organoids (Organoids_OM, black); GM Organoids (Organoids_GM, gray); WNT, Wingless MB (blue); SHH-A, Sonic Hedgehog MB-adulthood and childhood (red); SHH-B Sonic Hedgehog MB infant (dark red); G3, Group 3 MB (yellow); G4, Group 4 MB (green). f Hierarchical clustering and heatmap of beta values relative to the 39 high-quality CpG islands better discriminating MB subgroup in the Hovestadt set (Hovestadt et al.) 35 . The heatmap shows normalized methylation levels in organoid samples and MB samples. Clusters were obtained by means of Ward’s minimum variance method, using the Euclidean distance. Color legend: OM Organoids (Organoids_OM, black); GM Organoids (Organoids_ GM, gray); WNT, Wingless MB (blue); SHH-A, Sonic Hedgehog MB-adulthood and childhood (red); SHH-B Sonic Hedgehog MB infant (dark red); G3, Group 3 MB (yellow); G4, Group 4 MB (green). Scale bars 1 mm in ( b ), 100 µm in ( c , d ).
    Electroporation Machine, supplied by Amaxa, used in various techniques. Bioz Stars score: 86/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Amaxa electroporation nucleofection
    Cerebellar organoids <t>electroporation</t> with Otx2 / c-MYC induces Group 3 MB in vivo. a Schematic representation of in vivo injection of modified cerebellar organoids. b DAPI staining and GFP immunofluorescence (Venus) of the sagittal brain section of nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. c Confocal images of GFP (Venus) and PCNA immunofluorescence of tumors in nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. d NPR3 immunohistochemistry and Hematoxylin staining of tumor in nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. e MDS (multidimensional scaling) analysis performed on the 1000 most variable probes of the whole-genome DNA methylation data shows a close similarity between organoids and group 3 MBs. Color legend of the MDS plot as follows: OM Organoids (Organoids_OM, black); GM Organoids (Organoids_GM, gray); WNT, Wingless MB (blue); SHH-A, Sonic Hedgehog MB-adulthood and childhood (red); SHH-B Sonic Hedgehog MB infant (dark red); G3, Group 3 MB (yellow); G4, Group 4 MB (green). f Hierarchical clustering and heatmap of beta values relative to the 39 high-quality CpG islands better discriminating MB subgroup in the Hovestadt set (Hovestadt et al.) 35 . The heatmap shows normalized methylation levels in organoid samples and MB samples. Clusters were obtained by means of Ward’s minimum variance method, using the Euclidean distance. Color legend: OM Organoids (Organoids_OM, black); GM Organoids (Organoids_ GM, gray); WNT, Wingless MB (blue); SHH-A, Sonic Hedgehog MB-adulthood and childhood (red); SHH-B Sonic Hedgehog MB infant (dark red); G3, Group 3 MB (yellow); G4, Group 4 MB (green). Scale bars 1 mm in ( b ), 100 µm in ( c , d ).
    Electroporation Nucleofection, supplied by Amaxa, used in various techniques. Bioz Stars score: 86/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Amaxa electroporation system
    <t>Electroporation</t>
    Electroporation System, supplied by Amaxa, used in various techniques. Bioz Stars score: 92/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Bio-Rad electroporation system
    EBNA3C competes with p21 for Pim-1 binding. A–B) 10 million HEK-293 cells were transfected with different combinations of Myc-tagged Pim-1, Flag-tagged p21, untagged-EBNA3C expression vectors as indicated by <t>electroporation.</t> A) IP was performed with anti-Myc antibody and IP complexes were resolved by 10% SDS-PAGE. Western blot was performed with indicated antibodies. B) Above mentioned transfected cells were treated with MG132 drug for 6 hrs and protein lysates were prepared by RIPA buffer. Western blot analysis was performed by indicated antibodies.
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    Ichor Medical electroporation system
    EBNA3C competes with p21 for Pim-1 binding. A–B) 10 million HEK-293 cells were transfected with different combinations of Myc-tagged Pim-1, Flag-tagged p21, untagged-EBNA3C expression vectors as indicated by <t>electroporation.</t> A) IP was performed with anti-Myc antibody and IP complexes were resolved by 10% SDS-PAGE. Western blot was performed with indicated antibodies. B) Above mentioned transfected cells were treated with MG132 drug for 6 hrs and protein lysates were prepared by RIPA buffer. Western blot analysis was performed by indicated antibodies.
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    Bio-Rad electroporation tube
    EBNA3C competes with p21 for Pim-1 binding. A–B) 10 million HEK-293 cells were transfected with different combinations of Myc-tagged Pim-1, Flag-tagged p21, untagged-EBNA3C expression vectors as indicated by <t>electroporation.</t> A) IP was performed with anti-Myc antibody and IP complexes were resolved by 10% SDS-PAGE. Western blot was performed with indicated antibodies. B) Above mentioned transfected cells were treated with MG132 drug for 6 hrs and protein lysates were prepared by RIPA buffer. Western blot analysis was performed by indicated antibodies.
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    Lonza electroporation
    EBNA3C competes with p21 for Pim-1 binding. A–B) 10 million HEK-293 cells were transfected with different combinations of Myc-tagged Pim-1, Flag-tagged p21, untagged-EBNA3C expression vectors as indicated by <t>electroporation.</t> A) IP was performed with anti-Myc antibody and IP complexes were resolved by 10% SDS-PAGE. Western blot was performed with indicated antibodies. B) Above mentioned transfected cells were treated with MG132 drug for 6 hrs and protein lysates were prepared by RIPA buffer. Western blot analysis was performed by indicated antibodies.
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    Amaxa electroporation buffer
    Simplified, Efficient Genome Editing Using scCRISPR (A) Schematic shows the scCRISPR/Cas9 process that occurs inside target cells. (B) Histograms show flow cytometric GFP fluorescence (x axis) in Hist1h3a mouse ESCs (left) after <t>electroporation</t> with Cas9 and plasmid sgRNA (second from left), sgPal1 plasmid alone (third from left), and sgPal1 plasmid and sgGFP homology fragment with standard-length arms (fourth from left). (C) Fluorescence microscopy shows GFP fluorescence in Hist1h3a -GFP mouse ESCs (left) after targeting with Cas9 and plasmid sgRNA (second from left) and sgPal1 plasmid and sgGFP homology fragment (third from left). (D) Histograms show flow cytometric GFP fluorescence (x axis) in Hist1h3a -GFP knockin mouse ESCs after electroporation with Cas9 and (from left to right) sgPal1, sgPal7, and sgPal8 plasmids together with a long sgGFP homology fragment. (E) MiSeq plasmid copy numbers per cell of sgPal1, sgGFP, and the three most frequently mismatched sgGFP species 96 hr after co-electroporation of mouse ESCs are shown. (F) Multiplexed mutation of GFP (x axis) and dsRed (y axis) in Hist1h3a -GFP Rosa26 -dsRed mouse ESCs (left) after co-introduction of Cas9, sgPal1 plasmid, and sgGFP and sgDsRed long-armed homology fragments (right) is shown. See also Figure S1 .
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    MaxCyte electroporation buffer
    Similar signaling capacity and cytotocixity of mRNA electroporated and retrovirally transduced T cells . ( a ) Sensitivity of T-cell activation using T cells produced by mRNA <t>electroporation</t> compared with retroviral transduction. Results are displayed as percentage of maximum IFN-γ response obtained from intracellular cytokine staining. ( b ) Dose dependent lysis of HepG2-env targets (solid symbols and line) or HepG2-core targets (open symbols and dotted line) by electroporated T cells (triangle) compared with retrovirally transduced T cells (circle). Results are displayed as mean of triplicate measurements + SD. ( c ) Expression of perforin (left panel) and granzyme (right panel) on a representative activated T cells electroporated (red histograms) or retrovirally transduced (blue histograms) with s183-TCR, after 5 hours coculture with peptide-loaded T2 cells. Coculture with unpulsed T2 cells served as negative control (gray histograms). MFI of perforin and granzyme are indicated. ( d ) mRNA electroporated T cells have T CM -like phenotype. Phenotype of total lymphocytes (left panel), CD8 + pentamer + T cells (middle panel) and CD4 + Vb3 + T cells (right panel) in s183-TCR mRNA electroporated T cells (red shaded area) compared with retrovirally transduced T cells (blue shaded area). The percentages of CD45RA +/- and CD62L +/- cells within total lymphocytes or the gated CD8 + pentamer + or CD4 + Vb3 + populations were determined by FACS. Cells were classified into different subsets: naive (CD45RA + CD62L + ), T CM (CD45RA − CD62L + ), T EM (CD45RA − CD62L − ) and terminally differentiated EM (CD45RA + CD62L − ). Results expressed as mean + SD ( n = 3).
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    Teknova electroporation buffer
    Similar signaling capacity and cytotocixity of mRNA electroporated and retrovirally transduced T cells . ( a ) Sensitivity of T-cell activation using T cells produced by mRNA <t>electroporation</t> compared with retroviral transduction. Results are displayed as percentage of maximum IFN-γ response obtained from intracellular cytokine staining. ( b ) Dose dependent lysis of HepG2-env targets (solid symbols and line) or HepG2-core targets (open symbols and dotted line) by electroporated T cells (triangle) compared with retrovirally transduced T cells (circle). Results are displayed as mean of triplicate measurements + SD. ( c ) Expression of perforin (left panel) and granzyme (right panel) on a representative activated T cells electroporated (red histograms) or retrovirally transduced (blue histograms) with s183-TCR, after 5 hours coculture with peptide-loaded T2 cells. Coculture with unpulsed T2 cells served as negative control (gray histograms). MFI of perforin and granzyme are indicated. ( d ) mRNA electroporated T cells have T CM -like phenotype. Phenotype of total lymphocytes (left panel), CD8 + pentamer + T cells (middle panel) and CD4 + Vb3 + T cells (right panel) in s183-TCR mRNA electroporated T cells (red shaded area) compared with retrovirally transduced T cells (blue shaded area). The percentages of CD45RA +/- and CD62L +/- cells within total lymphocytes or the gated CD8 + pentamer + or CD4 + Vb3 + populations were determined by FACS. Cells were classified into different subsets: naive (CD45RA + CD62L + ), T CM (CD45RA − CD62L + ), T EM (CD45RA − CD62L − ) and terminally differentiated EM (CD45RA + CD62L − ). Results expressed as mean + SD ( n = 3).
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    Fisher Scientific electroporation cuvette
    The in vitro assessment IRE outcome on a non-cancerous cancer cell line (human dermal fibroblasts, HDFs) to irreversible <t>electroporation.</t> Viabilities were assessed at a range of electric field (500 to 1250 V/cm) and different number of pulses (10, 50 and 99). All data points are the result of n = 4 trials and are reported as the average ± standard deviation.
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    USA Scientific Inc electroporation cuvette
    Suppression of S. mansoni ABC transporter expression enhances responsiveness of adult worms to PZQ. Adult schistosomes were electroporated with siRNAs targeting luciferase (Control) or the S. mansoni ABC transporters SMDR2, SmMRP1, ABCA4, ABCB6, and MRP7/ABCC10 (KD), as described in Materials and Methods . Following <t>electroporation,</t> worms were incubated in schistosome medium for 2 days, and then sorted into 2–3 worm pairs per well in a 12-well plate. They were then incubated in medium alone plus 0.5% DMSO, or in medium plus 800 nM PZQ, as described in Fig. 1 , and subsequently analyzed for motility. n = 6 for the experiments without PZQ and n = 18 (KD) or 19 (Control) for the experiments in PZQ. Motility was measured as in Fig. 1 , and results for individual worms are plotted. ** indicates P
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    Harvard Bioscience electroporation cuvettes
    <t>Electroporation</t> of vertebrate and mosquito cell lines with EILV reporter chimeras. RNA was transcribed in vitro , each cell line was electroporated with ∼10 μg of RNA, and phase-contrast (left) and fluorescence (right) micrographs were taken at 1 and 4 days postelectroporation (dpe).
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    Harvard Bioscience electroporation generator
    Overexpression of Ascl1 changes the migratory behavior of neurons in the dorsal telencephalon. Ascl1 or US2 control expression construct were co-electroporated with a GFP expression plasmid. Two days after <t>electroporation</t> to the dorsal telencephalon, brains of E17.5 rats were dissected and sectioned in the coronal plane for slice culture and live imaging recording. The video from 5 to 11 hours after recording was used for tracking the migratory behavior. We set 0° to 180° axis in parallel to the lateral ventricle and the dorsomedial side as 180°. The line connecting the cell body location in the first frame (starting point) and the last frame (end point) of the video was used for measuring migratory angle. The average migratory rate was calculated as accumulated distance of every six-minute interval divided by recording time. 50 GFP-positive cells were counted for the control and 32 were counted for Ascl1 group. All counts were plotted into histograms according to their migratory angle or rate. ( A ) In the control group, most GFP-positive cells migrated radially in a migratory angle between 90° to 130°. ( B ) In Ascl1 group, GFP-positive cells were categorized into radially (90°–130°) and tangentially (140°–180°) migrating cells. ( C ) In the control group, GFP-positive cells migrated in the rate of 12.9 ± 3.6 μm/hour (mean ± SEM). ( D ) In Ascl1 group, radially migrating cells migrated in the rate of 30.2 ± 8.7 μm/hour and tangentially migrating cells migrated in the rate of 40.8 ± 9.3 μm/hour.
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    Image Search Results


    miR-195 and miR-497 decrease osteogenesis in human primary MSC A. miR-195 and miR-497 levels after electroporation of human MSC with miR-195 and miR-497 were determined by quantitative real time PCR. Small nuclear RNA U6 was used as reference gene. Values shown represent 2 independent experiments and are relative to scrambled negative control (SCR). B. ALP staining 7 days after electroporation of human MSC with scrambled negative control (SCR), miR-195 or miR-497 (5X; microscope scale: 50 μm; photography scale: 2 mm). C. ALP and RUNX2 expression levels in human MSC electroporated with either SCR, miR-195, miR-497. D. ALP expression levels 48h after MSC transfection with SCR negative control or anti-miR-195; ALP staining quantification in MSC transfected with SCR negative control or anti-miR-195 after 5 days of incubation with osteogenic supplements. Values shown represent 2 independent experiments and are relative to SCR (mean±SD; * P

    Journal: Oncotarget

    Article Title: miR-195 in human primary mesenchymal stromal/stem cells regulates proliferation, osteogenesis and paracrine effect on angiogenesis

    doi:

    Figure Lengend Snippet: miR-195 and miR-497 decrease osteogenesis in human primary MSC A. miR-195 and miR-497 levels after electroporation of human MSC with miR-195 and miR-497 were determined by quantitative real time PCR. Small nuclear RNA U6 was used as reference gene. Values shown represent 2 independent experiments and are relative to scrambled negative control (SCR). B. ALP staining 7 days after electroporation of human MSC with scrambled negative control (SCR), miR-195 or miR-497 (5X; microscope scale: 50 μm; photography scale: 2 mm). C. ALP and RUNX2 expression levels in human MSC electroporated with either SCR, miR-195, miR-497. D. ALP expression levels 48h after MSC transfection with SCR negative control or anti-miR-195; ALP staining quantification in MSC transfected with SCR negative control or anti-miR-195 after 5 days of incubation with osteogenic supplements. Values shown represent 2 independent experiments and are relative to SCR (mean±SD; * P

    Article Snippet: Electroporations and transfections Human primary MSC (Lonza) (0.5×106 ) were mixed with Pre-miR miRNA Precursors miR-195, Pre-miR miRNA Precursors miR-497 or Pre-miR miRNA Precursor Negative Control (Scrambled - SCR) (Life Technologies) in an electroporation cuvette and electroporated using OPTI-MEM I (Invitrogen, Life Technologies) in a Gene Pulser Xcell Electroporation Systems (Bio-Rad) with the following conditions: voltage - 250 V, capacitance - 950 μF, resistance - 200 Ω [ ].

    Techniques: Electroporation, Real-time Polymerase Chain Reaction, Negative Control, ALP Assay, Staining, Microscopy, Expressing, Transfection, Incubation

    miR-195 expression impairs MSC proliferation A. A representative experiment on cell viability of miR-195 and miR-497 in human MSC. Fluorescence of resorufin was measured every 24 hours for 4 days post-electroporation with SCR, miR-195 or miR-497. Values represent the mean±SD of 5 replicates (*** P

    Journal: Oncotarget

    Article Title: miR-195 in human primary mesenchymal stromal/stem cells regulates proliferation, osteogenesis and paracrine effect on angiogenesis

    doi:

    Figure Lengend Snippet: miR-195 expression impairs MSC proliferation A. A representative experiment on cell viability of miR-195 and miR-497 in human MSC. Fluorescence of resorufin was measured every 24 hours for 4 days post-electroporation with SCR, miR-195 or miR-497. Values represent the mean±SD of 5 replicates (*** P

    Article Snippet: Electroporations and transfections Human primary MSC (Lonza) (0.5×106 ) were mixed with Pre-miR miRNA Precursors miR-195, Pre-miR miRNA Precursors miR-497 or Pre-miR miRNA Precursor Negative Control (Scrambled - SCR) (Life Technologies) in an electroporation cuvette and electroporated using OPTI-MEM I (Invitrogen, Life Technologies) in a Gene Pulser Xcell Electroporation Systems (Bio-Rad) with the following conditions: voltage - 250 V, capacitance - 950 μF, resistance - 200 Ω [ ].

    Techniques: Expressing, Fluorescence, Electroporation

    Efficiency of electroporation and cell recovery using different methods. Naïve T-cells electroporated with an mRNA for the fluorescent protein Ruby. ( a ) Percentage of Ruby positive cells following electroporation using three different commercially available electroporators: BTX, Amaxa from Lonza and Neon from ThermoFisher. U-14, V-23 and T-23 are electroporation setting on the Amaxa machine. 1700 and 2150 are the voltages used on the Neon (see Materials and methods for more details). ( b ) Number of cell recovered following electroporation- a product of cell viability and cell count compared to starting condition (dashed grey line).

    Journal: eLife

    Article Title: A tissue-like platform for studying engineered quiescent human T-cells’ interactions with dendritic cells

    doi: 10.7554/eLife.48221

    Figure Lengend Snippet: Efficiency of electroporation and cell recovery using different methods. Naïve T-cells electroporated with an mRNA for the fluorescent protein Ruby. ( a ) Percentage of Ruby positive cells following electroporation using three different commercially available electroporators: BTX, Amaxa from Lonza and Neon from ThermoFisher. U-14, V-23 and T-23 are electroporation setting on the Amaxa machine. 1700 and 2150 are the voltages used on the Neon (see Materials and methods for more details). ( b ) Number of cell recovered following electroporation- a product of cell viability and cell count compared to starting condition (dashed grey line).

    Article Snippet: For Amaxa electroporation, the human T-cell nucleofector kit (VAPA-1002) was used and the manufacturer's protocol was followed.

    Techniques: Electroporation, Cell Counting

    Molecular ( a – c ) and biological ( d – f ) changes induced by Echinococcus granulosus Calmodulin (EgCaM)-specific siRNA using three delivery methods in different developmental stages in vitro . ( a – c ) EgCaM expression profile in different in vitro stages: protoscoleces (PSC) 3 and 8 days after electroporation (EP), soaking (SK) and electro-soaking (ES); microcysts (MC) and strobilated worms (SW). ( d ) Viability changes of protoscoleces treated with EgCaM-specific siRNA. ( e ) Size changes of 30 protoscoleces treated with EgCaM-specific siRNA. ( f ) Changes in body contractions per minute in the strobilated worms treated with EgCaM-specific siRNA. Data was compared to the controls: control protoscoleces (Ctrl) and negative siRNA control (siR-Ctrl). The gene expression data for siR-Ctrl were only demonstrated for electro-soaking method. Bars show the mean ± standard deviation (SD) derived from duplicates experiments. (**P

    Journal: Scientific Reports

    Article Title: Calmodulin-specific small interfering RNA induces consistent expression suppression and morphological changes in Echinococcus granulosus

    doi: 10.1038/s41598-019-40656-w

    Figure Lengend Snippet: Molecular ( a – c ) and biological ( d – f ) changes induced by Echinococcus granulosus Calmodulin (EgCaM)-specific siRNA using three delivery methods in different developmental stages in vitro . ( a – c ) EgCaM expression profile in different in vitro stages: protoscoleces (PSC) 3 and 8 days after electroporation (EP), soaking (SK) and electro-soaking (ES); microcysts (MC) and strobilated worms (SW). ( d ) Viability changes of protoscoleces treated with EgCaM-specific siRNA. ( e ) Size changes of 30 protoscoleces treated with EgCaM-specific siRNA. ( f ) Changes in body contractions per minute in the strobilated worms treated with EgCaM-specific siRNA. Data was compared to the controls: control protoscoleces (Ctrl) and negative siRNA control (siR-Ctrl). The gene expression data for siR-Ctrl were only demonstrated for electro-soaking method. Bars show the mean ± standard deviation (SD) derived from duplicates experiments. (**P

    Article Snippet: For electroporation, protoscoleces were removed from the culture, resuspended in 200 μl of electroporation buffer containing the fluorescently labeled siRNA to give a final concentration of 50 nM, transferred to a 2 mm gap cuvette, and were subjected to time constant protocol (125 V, 20 ms) using Gene Pulser II (Bio-Rad, CA) , , .

    Techniques: In Vitro, Expressing, Electroporation, Standard Deviation, Derivative Assay

    Morphological changes in Echinococcus granulosus treated with Calmodulin (EgCaM)-specific siRNA in vitro . Effect of EgCaM suppression on microcysts ( a ) and strobilated worms ( b ) using three delivery methods, electroporation (EP), soaking (SK), electro-soaking (ES) compared to the negative siRNA control (siR-Ctrl). Note the morphological changes (arrowheads) in the microcysts (outer layer irregularities and shrinkage, malformations and darkened edges) and the strobilated worms (morphological abnormalities and body swelling) compared to the controls. Scale bar = 200 μm.

    Journal: Scientific Reports

    Article Title: Calmodulin-specific small interfering RNA induces consistent expression suppression and morphological changes in Echinococcus granulosus

    doi: 10.1038/s41598-019-40656-w

    Figure Lengend Snippet: Morphological changes in Echinococcus granulosus treated with Calmodulin (EgCaM)-specific siRNA in vitro . Effect of EgCaM suppression on microcysts ( a ) and strobilated worms ( b ) using three delivery methods, electroporation (EP), soaking (SK), electro-soaking (ES) compared to the negative siRNA control (siR-Ctrl). Note the morphological changes (arrowheads) in the microcysts (outer layer irregularities and shrinkage, malformations and darkened edges) and the strobilated worms (morphological abnormalities and body swelling) compared to the controls. Scale bar = 200 μm.

    Article Snippet: For electroporation, protoscoleces were removed from the culture, resuspended in 200 μl of electroporation buffer containing the fluorescently labeled siRNA to give a final concentration of 50 nM, transferred to a 2 mm gap cuvette, and were subjected to time constant protocol (125 V, 20 ms) using Gene Pulser II (Bio-Rad, CA) , , .

    Techniques: In Vitro, Electroporation

    GFP and VEGF gene expression of electroporated BMMSCs. Green fluorescence represents the successful electroporation of plasmid into BMMSCs. Electroporated cells after 48 h ( a and b ). G418 selection after 20 days ( d and e ) ( bright vision

    Journal: Cytotechnology

    Article Title: VEGF165 expressing bone marrow mesenchymal stem cells differentiate into hepatocytes under HGF and EGF induction in vitro

    doi: 10.1007/s10616-012-9439-0

    Figure Lengend Snippet: GFP and VEGF gene expression of electroporated BMMSCs. Green fluorescence represents the successful electroporation of plasmid into BMMSCs. Electroporated cells after 48 h ( a and b ). G418 selection after 20 days ( d and e ) ( bright vision

    Article Snippet: The 3rd generation BMMSCs were harvested and resuspended in electroporation buffer [containing 750 μl Opti-DMEM (Gibco, USA) without serum and 50 μg VEGF165 -pCMV6-AC-GFP plasmid (OriGene, Rockville/MD, USA) or pCMV6-AC-GFP plasmid (OriGene, USA)].

    Techniques: Expressing, Fluorescence, Electroporation, Plasmid Preparation, Selection

    Robo/Dll1 Signaling Regulates the Balance between Direct Neurogenesis and IPC Abundance in Chick Dorsal Pallium (A–D) Analysis of chick dorsal pallium at 6 days post-ovoposition (dpo), showing many neurons in the VZ (solid arrowheads) in the medial part (mDP; B) and basal mitoses (open arrowheads) in the lateral (lDP; C; n = 3 embryos; t tests). (E–H) ISH in chick DP at 6 dpo, and quantifications of intensity (au, arbitrary units). High magnifications show chkRobo1 (E’ and E”) and chkDll1 (G’ and G”) in the indicated regions. Panel shown in (G) is a tiled image. (I) qPCR analysis in the VZ of the regions and species indicated. Values are ratio Robo1 to Gapdh (n = 12–15 replicates; paired or independent samples t tests). (J–O) Experimental design to manipulate in ovo Robo and Dll1 in mDP and lDP, representative examples and quantifications of neurons in the VZ (Tbr1+) and basal mitoses (PH3, open arrowheads; n = 3-5 embryos per group; t tests in L and Q; one-way ANOVA followed by t tests in M and O). Red shadowing and asterisks in (L) and (O) indicate values within group corresponding to GFP− cells. (P and Q) Expression of Tbr2 in basal PH3+ mitoses (solid arrowhead, Tbr2+; open arrowhead, Tbr2−) upon electroporation of dnR1/2+Dll1 in mDP (as in J and K), and quantification (n = 4–5 embryos; t tests). (R–U) Analysis of neuronal clones (GFP+RFP+Tuj1+) upon overexpression of dnR1/2+Dll1+Gfp, representative examples and quantification (n = 52 clones Gfp, 59 clones dnR1/2+Dll1+Gfp, 3–7 embryos; t test or χ2-test). Values are mean + SEM; ∗ p

    Journal: Cell

    Article Title: Evolution of Cortical Neurogenesis in Amniotes Controlled by Robo Signaling Levels

    doi: 10.1016/j.cell.2018.06.007

    Figure Lengend Snippet: Robo/Dll1 Signaling Regulates the Balance between Direct Neurogenesis and IPC Abundance in Chick Dorsal Pallium (A–D) Analysis of chick dorsal pallium at 6 days post-ovoposition (dpo), showing many neurons in the VZ (solid arrowheads) in the medial part (mDP; B) and basal mitoses (open arrowheads) in the lateral (lDP; C; n = 3 embryos; t tests). (E–H) ISH in chick DP at 6 dpo, and quantifications of intensity (au, arbitrary units). High magnifications show chkRobo1 (E’ and E”) and chkDll1 (G’ and G”) in the indicated regions. Panel shown in (G) is a tiled image. (I) qPCR analysis in the VZ of the regions and species indicated. Values are ratio Robo1 to Gapdh (n = 12–15 replicates; paired or independent samples t tests). (J–O) Experimental design to manipulate in ovo Robo and Dll1 in mDP and lDP, representative examples and quantifications of neurons in the VZ (Tbr1+) and basal mitoses (PH3, open arrowheads; n = 3-5 embryos per group; t tests in L and Q; one-way ANOVA followed by t tests in M and O). Red shadowing and asterisks in (L) and (O) indicate values within group corresponding to GFP− cells. (P and Q) Expression of Tbr2 in basal PH3+ mitoses (solid arrowhead, Tbr2+; open arrowhead, Tbr2−) upon electroporation of dnR1/2+Dll1 in mDP (as in J and K), and quantification (n = 4–5 embryos; t tests). (R–U) Analysis of neuronal clones (GFP+RFP+Tuj1+) upon overexpression of dnR1/2+Dll1+Gfp, representative examples and quantification (n = 52 clones Gfp, 59 clones dnR1/2+Dll1+Gfp, 3–7 embryos; t test or χ2-test). Values are mean + SEM; ∗ p

    Article Snippet: During the electroporation cerebral organoids were placed in an electroporation chamber (Harvard Apparatus, Holliston, MA, USA) under a stereoscope and using a glass microcapillary 1-2 μL of plasmid DNAs was injected together with Fast Green (0.1%, Sigma) into different ventricles of the organoids.

    Techniques: In Situ Hybridization, Real-time Polymerase Chain Reaction, In Ovo, Expressing, Electroporation, Clone Assay, Over Expression

    Electric field enhancement by CNT in electroporation. (a) Sketch of electroporation with CNT-free (left) and CNT associated cells (right). Table insert: theoretical electric field ( E ) enhancement over the applied field ( E 0 ) by a nanotube of 1 μ m long. d denotes the distance to nanotube tip. a 1 and a 2 are the aspect ratios (i.e. diameter/length) of nanotubes, and equal to 0.1 and 0.5 respectively. (b) Electroporation time constants versus the voltages. A series of time constants of the CNT associated Bal17 cells were obtained under various voltages. They were compared to the time constant of regular Bal17 cells at 230 V. (c) Improvement of EGFP transfection by CNT mediated electroporation in Bal17 cells. The gates of EGFP positive cells were adjusted based on the Neg Contl and Pos Contl . The mean values of EGFP levels (left axis) and the percentages (right axis) of gated viable cells are shown. Neg Contl : negative control, cells speared and electroporated without plasmid; Pos Contl : positive control, cells without spearing and electroporated at 230 V with EGFP plasmid; 180, 200 and 220 V: CNT associated cells and electroporated with EGFP plasmids at corresponding voltages.

    Journal: Nanotechnology

    Article Title: Interaction between carbon nanotubes and mammalian cells: characterization by flow cytometry and application

    doi: 10.1088/0957-4484/19/34/345102

    Figure Lengend Snippet: Electric field enhancement by CNT in electroporation. (a) Sketch of electroporation with CNT-free (left) and CNT associated cells (right). Table insert: theoretical electric field ( E ) enhancement over the applied field ( E 0 ) by a nanotube of 1 μ m long. d denotes the distance to nanotube tip. a 1 and a 2 are the aspect ratios (i.e. diameter/length) of nanotubes, and equal to 0.1 and 0.5 respectively. (b) Electroporation time constants versus the voltages. A series of time constants of the CNT associated Bal17 cells were obtained under various voltages. They were compared to the time constant of regular Bal17 cells at 230 V. (c) Improvement of EGFP transfection by CNT mediated electroporation in Bal17 cells. The gates of EGFP positive cells were adjusted based on the Neg Contl and Pos Contl . The mean values of EGFP levels (left axis) and the percentages (right axis) of gated viable cells are shown. Neg Contl : negative control, cells speared and electroporated without plasmid; Pos Contl : positive control, cells without spearing and electroporated at 230 V with EGFP plasmid; 180, 200 and 220 V: CNT associated cells and electroporated with EGFP plasmids at corresponding voltages.

    Article Snippet: Each electroporation cuvette (cooled on ice) was loaded with 3 × 106 regular or CNT associated Bal17 cells in suspension adjusted to 200 μ l. A Gene Pulser electroporation system (BIO-RAD, Hercules, CA) was High Cap, 960 μ F. Voltages were selected from 100 to 250 V. For the transfection experiments, pEGFP plasmid DNA (Clontech, Mountain View, CA) was supplemented to the cell suspensions in the cuvettes.

    Techniques: Electroporation, Transfection, Negative Control, Plasmid Preparation, Positive Control

    NHEJ of a DSB model substrate in HeLa cells in vivo . Substrate XM was introduced into HeLa cells by electroporation. After transfection, cells were cultured for the indicated times after which substrate DNA was recovered and transfected into E.coli to assess formation of circular monomeric end joining products as described in Materials and Methods. NHEJ activity after 2 h in culture was taken as 100%. Error bars represent the standard deviation of two independent experiments.

    Journal: Nucleic Acids Research

    Article Title: The role of DNA polymerase activity in human non-homologous end joining

    doi:

    Figure Lengend Snippet: NHEJ of a DSB model substrate in HeLa cells in vivo . Substrate XM was introduced into HeLa cells by electroporation. After transfection, cells were cultured for the indicated times after which substrate DNA was recovered and transfected into E.coli to assess formation of circular monomeric end joining products as described in Materials and Methods. NHEJ activity after 2 h in culture was taken as 100%. Error bars represent the standard deviation of two independent experiments.

    Article Snippet: The cells were resuspended at 5 × 107 cells in the same medium and 400 µl aliquots were mixed with 1 µg DSB substrate and 50–100 ng pBluescript II SK+ (Stratagene) and transferred to electroporation cuvettes with a 4 mm gap (Bio-Rad).

    Techniques: Non-Homologous End Joining, In Vivo, Electroporation, Transfection, Cell Culture, Activity Assay, Standard Deviation

    CRF protein homo- and heterodimerization. (A) CRF proteins CRF1–CRF8 and just the CRF domain of CRF2 (otherwise noted as CRFD) were analysed for potential homo- and heterodimerization using both BiFC and Y2H. Positive interactions are noted as a (+), (+*) for both BiFC and Y2H together, and (–) for non-interactions. (B) A cartoon of the conserved regions of CRF1–CRF6 proteins, the highly related CRF7 and CRF8 proteins (lacking the C-terminal third of the protein, but containing both the CRF domain and the AP2/ERF domain), and CRFD (just the CRF domain of CRF2). (C, D) Representative examples of positive CRF protein homo- and heterodimerization by BiFC as indicated in A are shown both under UV light in the presence of Hoechst 33342 dye denoting the nucleus and using a YFP wavelength filter to visualize BiFC interaction. Additionally, representative examples of empty vector (EV) controls for both N- and C-terminal BiFC vectors (EVn and EVc) examined in A are shown versus various CRFs and each other. (C) BiFC interactions in protoplasts transformed via electroporation. (D) BiFC interactions in tobacco leaves transformed via Agrobacterium .

    Journal: Journal of Experimental Botany

    Article Title: CRFs form protein-protein interactions with each other and with members of the cytokinin signalling pathway in Arabidopsis via the CRF domain

    doi: 10.1093/jxb/err199

    Figure Lengend Snippet: CRF protein homo- and heterodimerization. (A) CRF proteins CRF1–CRF8 and just the CRF domain of CRF2 (otherwise noted as CRFD) were analysed for potential homo- and heterodimerization using both BiFC and Y2H. Positive interactions are noted as a (+), (+*) for both BiFC and Y2H together, and (–) for non-interactions. (B) A cartoon of the conserved regions of CRF1–CRF6 proteins, the highly related CRF7 and CRF8 proteins (lacking the C-terminal third of the protein, but containing both the CRF domain and the AP2/ERF domain), and CRFD (just the CRF domain of CRF2). (C, D) Representative examples of positive CRF protein homo- and heterodimerization by BiFC as indicated in A are shown both under UV light in the presence of Hoechst 33342 dye denoting the nucleus and using a YFP wavelength filter to visualize BiFC interaction. Additionally, representative examples of empty vector (EV) controls for both N- and C-terminal BiFC vectors (EVn and EVc) examined in A are shown versus various CRFs and each other. (C) BiFC interactions in protoplasts transformed via electroporation. (D) BiFC interactions in tobacco leaves transformed via Agrobacterium .

    Article Snippet: Protoplasts were centrifuged again, washing buffer was removed, and then protoplasts were resuspended in a final volume of washing/transformation buffer prior to transformation and placed on ice until transformation, usually within 1 h. A 100 μl aliquot of transformation buffer containing ∼105 protoplasts along with ∼40–50 μg of plasmid DNA for each plasmid used were given two rapid pulses of 300 V for electroporation in a 0.1 mm electroporation cuvette using an Eppendorf Electroporator 2510.

    Techniques: Bimolecular Fluorescence Complementation Assay, Plasmid Preparation, Transformation Assay, Electroporation

    CRF protein interactions with members of the cytokinin signalling pathway. (A) CRF proteins, CRF1–CRF8, and CRFD (C-terminal) were analysed for potential interactions with TCS pathway proteins (N-terminal) using BiFC. Positive interactions are noted as (+) and non-interactions as (–). (B, C) Representative examples of positive and negative CRF protein interactions with TCS proteins as well as an empty vector control (EVc: C-terminal) by BiFC as indicated in A are shown both under UV light in the presence of Hoechst 33342 dye denoting the nucleus and using a YFP wavelength filter to visualize BiFC interaction. (B) BiFC interactions in protoplasts transformed via electroporation. (C) BiFC interactions in tobacco leaves transformed via Agrobacterium .

    Journal: Journal of Experimental Botany

    Article Title: CRFs form protein-protein interactions with each other and with members of the cytokinin signalling pathway in Arabidopsis via the CRF domain

    doi: 10.1093/jxb/err199

    Figure Lengend Snippet: CRF protein interactions with members of the cytokinin signalling pathway. (A) CRF proteins, CRF1–CRF8, and CRFD (C-terminal) were analysed for potential interactions with TCS pathway proteins (N-terminal) using BiFC. Positive interactions are noted as (+) and non-interactions as (–). (B, C) Representative examples of positive and negative CRF protein interactions with TCS proteins as well as an empty vector control (EVc: C-terminal) by BiFC as indicated in A are shown both under UV light in the presence of Hoechst 33342 dye denoting the nucleus and using a YFP wavelength filter to visualize BiFC interaction. (B) BiFC interactions in protoplasts transformed via electroporation. (C) BiFC interactions in tobacco leaves transformed via Agrobacterium .

    Article Snippet: Protoplasts were centrifuged again, washing buffer was removed, and then protoplasts were resuspended in a final volume of washing/transformation buffer prior to transformation and placed on ice until transformation, usually within 1 h. A 100 μl aliquot of transformation buffer containing ∼105 protoplasts along with ∼40–50 μg of plasmid DNA for each plasmid used were given two rapid pulses of 300 V for electroporation in a 0.1 mm electroporation cuvette using an Eppendorf Electroporator 2510.

    Techniques: Bimolecular Fluorescence Complementation Assay, Plasmid Preparation, Transformation Assay, Electroporation

    Expression and permeabilization of cells. a CHO cells to set 1 parameters in 6-h time gap between EP and nsEP, observed 24 h after nsEP. Phase contrast image and fluorescence image of GFP expression (40× objective). b Permeabilization of cells observed by entry of propidium iodide, contrast phase and fluorescence image for classical electroporation protocol only of set 1 (100× objective). c – e Permeabilization of cells submitted to 200 nsEP only of sets 1, 2 and 3, respectively—phase contrast image and fluorescence-associated (100× objective)

    Journal: The Journal of Membrane Biology

    Article Title: Nanosecond Electric Pulse Effects on Gene Expression

    doi: 10.1007/s00232-013-9579-y

    Figure Lengend Snippet: Expression and permeabilization of cells. a CHO cells to set 1 parameters in 6-h time gap between EP and nsEP, observed 24 h after nsEP. Phase contrast image and fluorescence image of GFP expression (40× objective). b Permeabilization of cells observed by entry of propidium iodide, contrast phase and fluorescence image for classical electroporation protocol only of set 1 (100× objective). c – e Permeabilization of cells submitted to 200 nsEP only of sets 1, 2 and 3, respectively—phase contrast image and fluorescence-associated (100× objective)

    Article Snippet: Cell suspension (70 μl) was placed in electroporation cuvettes with built-in aluminum electrodes with a 1-mm gap (Eppendorf).

    Techniques: Expressing, Fluorescence, Electroporation

    Cerebellar organoids electroporation with Otx2 / c-MYC induces Group 3 MB in vivo. a Schematic representation of in vivo injection of modified cerebellar organoids. b DAPI staining and GFP immunofluorescence (Venus) of the sagittal brain section of nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. c Confocal images of GFP (Venus) and PCNA immunofluorescence of tumors in nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. d NPR3 immunohistochemistry and Hematoxylin staining of tumor in nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. e MDS (multidimensional scaling) analysis performed on the 1000 most variable probes of the whole-genome DNA methylation data shows a close similarity between organoids and group 3 MBs. Color legend of the MDS plot as follows: OM Organoids (Organoids_OM, black); GM Organoids (Organoids_GM, gray); WNT, Wingless MB (blue); SHH-A, Sonic Hedgehog MB-adulthood and childhood (red); SHH-B Sonic Hedgehog MB infant (dark red); G3, Group 3 MB (yellow); G4, Group 4 MB (green). f Hierarchical clustering and heatmap of beta values relative to the 39 high-quality CpG islands better discriminating MB subgroup in the Hovestadt set (Hovestadt et al.) 35 . The heatmap shows normalized methylation levels in organoid samples and MB samples. Clusters were obtained by means of Ward’s minimum variance method, using the Euclidean distance. Color legend: OM Organoids (Organoids_OM, black); GM Organoids (Organoids_ GM, gray); WNT, Wingless MB (blue); SHH-A, Sonic Hedgehog MB-adulthood and childhood (red); SHH-B Sonic Hedgehog MB infant (dark red); G3, Group 3 MB (yellow); G4, Group 4 MB (green). Scale bars 1 mm in ( b ), 100 µm in ( c , d ).

    Journal: Nature Communications

    Article Title: Modeling medulloblastoma in vivo and with human cerebellar organoids

    doi: 10.1038/s41467-019-13989-3

    Figure Lengend Snippet: Cerebellar organoids electroporation with Otx2 / c-MYC induces Group 3 MB in vivo. a Schematic representation of in vivo injection of modified cerebellar organoids. b DAPI staining and GFP immunofluorescence (Venus) of the sagittal brain section of nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. c Confocal images of GFP (Venus) and PCNA immunofluorescence of tumors in nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. d NPR3 immunohistochemistry and Hematoxylin staining of tumor in nude mouse 1 month after injection of human cerebellar organoids electroporated with pPBase + pPBMyc + pPBOtx2 + pPBVenus. e MDS (multidimensional scaling) analysis performed on the 1000 most variable probes of the whole-genome DNA methylation data shows a close similarity between organoids and group 3 MBs. Color legend of the MDS plot as follows: OM Organoids (Organoids_OM, black); GM Organoids (Organoids_GM, gray); WNT, Wingless MB (blue); SHH-A, Sonic Hedgehog MB-adulthood and childhood (red); SHH-B Sonic Hedgehog MB infant (dark red); G3, Group 3 MB (yellow); G4, Group 4 MB (green). f Hierarchical clustering and heatmap of beta values relative to the 39 high-quality CpG islands better discriminating MB subgroup in the Hovestadt set (Hovestadt et al.) 35 . The heatmap shows normalized methylation levels in organoid samples and MB samples. Clusters were obtained by means of Ward’s minimum variance method, using the Euclidean distance. Color legend: OM Organoids (Organoids_OM, black); GM Organoids (Organoids_ GM, gray); WNT, Wingless MB (blue); SHH-A, Sonic Hedgehog MB-adulthood and childhood (red); SHH-B Sonic Hedgehog MB infant (dark red); G3, Group 3 MB (yellow); G4, Group 4 MB (green). Scale bars 1 mm in ( b ), 100 µm in ( c , d ).

    Article Snippet: Organoids were transferred inside the Electroporation cuvettes (VWR, ECN 732-1136, 2 mm), and electroporation was performed with the Gene Pulser XcellTM.

    Techniques: Electroporation, In Vivo, Injection, Modification, Staining, Immunofluorescence, Immunohistochemistry, DNA Methylation Assay, Methylation

    Cerebellar organoids electroporation with Gfi1 / c-MYC and Otx2 / c-MYC induces overproliferation. a Schematic representation of organoids electroporation. b Brightfield and fluorescence images of cerebellar organoids at day 60 electroporated at day 35 with pPBVenus. c Brightfield and fluorescence images of cerebellar organoids at day 60 electroporated at day 35 with pPBase + pPBMyc + pPBGfi1 + pPBVenus. d Brightfield and fluorescence images of cerebellar organoids at day 60 electroporated at day 35 with pPBase + pPBMyc + pPBOtx2 + pPBVenus. Arrows in ( b , c , d ) indicate Venus-positive cells. e Confocal images of GFP (Venus) and PCNA immunofluorescence of cerebellar organoids at day 60 electroporated at day 35 with pPBase + pPBVenus + pPBMyc and pPBOtx2. Arrows indicate double-positive cells. f Confocal images of GFP (Venus) and β3-tubulin immunofluorescence of cerebellar organoids at day 60 electroporated at day 35 with pPBase + pPBMyc + pPBOtx2 + pPBVenus. Arrows indicate β3-tubulin-negative cells. g Quantification of cerebellar organoids at day 60, electroporated at day 35 with either pPBVenus or pPBase + pPBMyc + pPBOtx2 + pPBVenus (OM) or pPBase + pPBMyc + pPBGfi1 + pPBVenus (GM). n = 6 biologically independent organoids. h Quantification of cerebellar organoids at day 40, electroporated at day 35 with either pPBVenus or pPBase + pPBMyc + pPBOtx2 + pPBVenus (OM) or pPBase + pPBMyc + pPBGfi1 + pPBVenus (GM). n = 6 biologically independent organoids. Error bars in ( g , e ) represent standard error of the mean. Scale bars 250 µm in ( b – d ),100 µm in ( e , f ). Paired Student's t test, two tails. * p -value

    Journal: Nature Communications

    Article Title: Modeling medulloblastoma in vivo and with human cerebellar organoids

    doi: 10.1038/s41467-019-13989-3

    Figure Lengend Snippet: Cerebellar organoids electroporation with Gfi1 / c-MYC and Otx2 / c-MYC induces overproliferation. a Schematic representation of organoids electroporation. b Brightfield and fluorescence images of cerebellar organoids at day 60 electroporated at day 35 with pPBVenus. c Brightfield and fluorescence images of cerebellar organoids at day 60 electroporated at day 35 with pPBase + pPBMyc + pPBGfi1 + pPBVenus. d Brightfield and fluorescence images of cerebellar organoids at day 60 electroporated at day 35 with pPBase + pPBMyc + pPBOtx2 + pPBVenus. Arrows in ( b , c , d ) indicate Venus-positive cells. e Confocal images of GFP (Venus) and PCNA immunofluorescence of cerebellar organoids at day 60 electroporated at day 35 with pPBase + pPBVenus + pPBMyc and pPBOtx2. Arrows indicate double-positive cells. f Confocal images of GFP (Venus) and β3-tubulin immunofluorescence of cerebellar organoids at day 60 electroporated at day 35 with pPBase + pPBMyc + pPBOtx2 + pPBVenus. Arrows indicate β3-tubulin-negative cells. g Quantification of cerebellar organoids at day 60, electroporated at day 35 with either pPBVenus or pPBase + pPBMyc + pPBOtx2 + pPBVenus (OM) or pPBase + pPBMyc + pPBGfi1 + pPBVenus (GM). n = 6 biologically independent organoids. h Quantification of cerebellar organoids at day 40, electroporated at day 35 with either pPBVenus or pPBase + pPBMyc + pPBOtx2 + pPBVenus (OM) or pPBase + pPBMyc + pPBGfi1 + pPBVenus (GM). n = 6 biologically independent organoids. Error bars in ( g , e ) represent standard error of the mean. Scale bars 250 µm in ( b – d ),100 µm in ( e , f ). Paired Student's t test, two tails. * p -value

    Article Snippet: Organoids were transferred inside the Electroporation cuvettes (VWR, ECN 732-1136, 2 mm), and electroporation was performed with the Gene Pulser XcellTM.

    Techniques: Electroporation, Fluorescence, Immunofluorescence

    ( a and b ) The normalized fluorescence ( NF ) of intracellular Fluorescein-Dextran as a result of double-pulse electroporation. ( Symbols ) Experimental data; ( curves ) least-square fitting. For all cases, the first pulse was always E 1 = 100,000 V/m and t 1 = 0.001s. To see this figure in color, go online.

    Journal: Biophysical Journal

    Article Title: Scaling Relationship and Optimization of Double-Pulse Electroporation

    doi: 10.1016/j.bpj.2013.12.045

    Figure Lengend Snippet: ( a and b ) The normalized fluorescence ( NF ) of intracellular Fluorescein-Dextran as a result of double-pulse electroporation. ( Symbols ) Experimental data; ( curves ) least-square fitting. For all cases, the first pulse was always E 1 = 100,000 V/m and t 1 = 0.001s. To see this figure in color, go online.

    Article Snippet: A volume of 90 μ L of cell suspension was placed into an electroporation cuvette (model No. 89047-206; VWR, Philadelphia, PA).

    Techniques: Fluorescence, Electroporation

    The percentage of viable cells after double-pulse electroporation. ( Symbols ) Experimental data; ( curves ) least-square fitting. For all cases, the first pulse was always E 1 = 100,000 V/m and t 1 = 0.001 s. To see this figure in color, go online.

    Journal: Biophysical Journal

    Article Title: Scaling Relationship and Optimization of Double-Pulse Electroporation

    doi: 10.1016/j.bpj.2013.12.045

    Figure Lengend Snippet: The percentage of viable cells after double-pulse electroporation. ( Symbols ) Experimental data; ( curves ) least-square fitting. For all cases, the first pulse was always E 1 = 100,000 V/m and t 1 = 0.001 s. To see this figure in color, go online.

    Article Snippet: A volume of 90 μ L of cell suspension was placed into an electroporation cuvette (model No. 89047-206; VWR, Philadelphia, PA).

    Techniques: Electroporation

    Electroporation

    Journal: Annals of biomedical engineering

    Article Title: Physical non-viral gene delivery methods for tissue engineering

    doi: 10.1007/s10439-012-0678-1

    Figure Lengend Snippet: Electroporation

    Article Snippet: Nucleofection™ is a patented commercial electroporation system created by Amaxa, and owned by Lonza.

    Techniques: Electroporation

    EBNA3C competes with p21 for Pim-1 binding. A–B) 10 million HEK-293 cells were transfected with different combinations of Myc-tagged Pim-1, Flag-tagged p21, untagged-EBNA3C expression vectors as indicated by electroporation. A) IP was performed with anti-Myc antibody and IP complexes were resolved by 10% SDS-PAGE. Western blot was performed with indicated antibodies. B) Above mentioned transfected cells were treated with MG132 drug for 6 hrs and protein lysates were prepared by RIPA buffer. Western blot analysis was performed by indicated antibodies.

    Journal: PLoS Pathogens

    Article Title: EBNA3C Augments Pim-1 Mediated Phosphorylation and Degradation of p21 to Promote B-Cell Proliferation

    doi: 10.1371/journal.ppat.1004304

    Figure Lengend Snippet: EBNA3C competes with p21 for Pim-1 binding. A–B) 10 million HEK-293 cells were transfected with different combinations of Myc-tagged Pim-1, Flag-tagged p21, untagged-EBNA3C expression vectors as indicated by electroporation. A) IP was performed with anti-Myc antibody and IP complexes were resolved by 10% SDS-PAGE. Western blot was performed with indicated antibodies. B) Above mentioned transfected cells were treated with MG132 drug for 6 hrs and protein lysates were prepared by RIPA buffer. Western blot analysis was performed by indicated antibodies.

    Article Snippet: Transfections Transfection in HEK-293, MEF and B-cells were performed by electroporation system with Bio-Rad Gene Pulser II electroporator.

    Techniques: Binding Assay, Transfection, Expressing, Electroporation, SDS Page, Western Blot

    EBNA3C destabilizes p21 through Pim-1 independent of the DNA damage response. A) 10 million HEK-293 cells were transfected with combinations of Myc-tagged Pim-1, Flag-tagged p21, untagged-EBNA3C expression by electroporation. After 36 hrs of post-transfection, cells were treated with cyclohexamide for indicated time points in DMEM medium containing either serum/DMSO or 0.1% FBS with 5 µM etoposide. Protein samples were resolved by 10% SDS-PAGE and Western blots were performed with indicated antibodies. B–C) 50 million EBV negative BJAB cells, EBNA3C expressing BJAB10, EBV transformed LCL1, sh-Ctrl, sh-E3C LCL1 cells were incubated with cyclohexamide for specific time points in RPMI medium containing serum/DMSO or 0.1% FBS with 5 µM etoposide. Samples were resolved by 10% SDS-PAGE and Western blot analysis was performed with specific antibodies.

    Journal: PLoS Pathogens

    Article Title: EBNA3C Augments Pim-1 Mediated Phosphorylation and Degradation of p21 to Promote B-Cell Proliferation

    doi: 10.1371/journal.ppat.1004304

    Figure Lengend Snippet: EBNA3C destabilizes p21 through Pim-1 independent of the DNA damage response. A) 10 million HEK-293 cells were transfected with combinations of Myc-tagged Pim-1, Flag-tagged p21, untagged-EBNA3C expression by electroporation. After 36 hrs of post-transfection, cells were treated with cyclohexamide for indicated time points in DMEM medium containing either serum/DMSO or 0.1% FBS with 5 µM etoposide. Protein samples were resolved by 10% SDS-PAGE and Western blots were performed with indicated antibodies. B–C) 50 million EBV negative BJAB cells, EBNA3C expressing BJAB10, EBV transformed LCL1, sh-Ctrl, sh-E3C LCL1 cells were incubated with cyclohexamide for specific time points in RPMI medium containing serum/DMSO or 0.1% FBS with 5 µM etoposide. Samples were resolved by 10% SDS-PAGE and Western blot analysis was performed with specific antibodies.

    Article Snippet: Transfections Transfection in HEK-293, MEF and B-cells were performed by electroporation system with Bio-Rad Gene Pulser II electroporator.

    Techniques: Transfection, Expressing, Electroporation, SDS Page, Western Blot, Transformation Assay, Incubation

    Simplified, Efficient Genome Editing Using scCRISPR (A) Schematic shows the scCRISPR/Cas9 process that occurs inside target cells. (B) Histograms show flow cytometric GFP fluorescence (x axis) in Hist1h3a mouse ESCs (left) after electroporation with Cas9 and plasmid sgRNA (second from left), sgPal1 plasmid alone (third from left), and sgPal1 plasmid and sgGFP homology fragment with standard-length arms (fourth from left). (C) Fluorescence microscopy shows GFP fluorescence in Hist1h3a -GFP mouse ESCs (left) after targeting with Cas9 and plasmid sgRNA (second from left) and sgPal1 plasmid and sgGFP homology fragment (third from left). (D) Histograms show flow cytometric GFP fluorescence (x axis) in Hist1h3a -GFP knockin mouse ESCs after electroporation with Cas9 and (from left to right) sgPal1, sgPal7, and sgPal8 plasmids together with a long sgGFP homology fragment. (E) MiSeq plasmid copy numbers per cell of sgPal1, sgGFP, and the three most frequently mismatched sgGFP species 96 hr after co-electroporation of mouse ESCs are shown. (F) Multiplexed mutation of GFP (x axis) and dsRed (y axis) in Hist1h3a -GFP Rosa26 -dsRed mouse ESCs (left) after co-introduction of Cas9, sgPal1 plasmid, and sgGFP and sgDsRed long-armed homology fragments (right) is shown. See also Figure S1 .

    Journal: Stem Cell Reports

    Article Title: Cloning-free CRISPR

    doi: 10.1016/j.stemcr.2015.09.022

    Figure Lengend Snippet: Simplified, Efficient Genome Editing Using scCRISPR (A) Schematic shows the scCRISPR/Cas9 process that occurs inside target cells. (B) Histograms show flow cytometric GFP fluorescence (x axis) in Hist1h3a mouse ESCs (left) after electroporation with Cas9 and plasmid sgRNA (second from left), sgPal1 plasmid alone (third from left), and sgPal1 plasmid and sgGFP homology fragment with standard-length arms (fourth from left). (C) Fluorescence microscopy shows GFP fluorescence in Hist1h3a -GFP mouse ESCs (left) after targeting with Cas9 and plasmid sgRNA (second from left) and sgPal1 plasmid and sgGFP homology fragment (third from left). (D) Histograms show flow cytometric GFP fluorescence (x axis) in Hist1h3a -GFP knockin mouse ESCs after electroporation with Cas9 and (from left to right) sgPal1, sgPal7, and sgPal8 plasmids together with a long sgGFP homology fragment. (E) MiSeq plasmid copy numbers per cell of sgPal1, sgGFP, and the three most frequently mismatched sgGFP species 96 hr after co-electroporation of mouse ESCs are shown. (F) Multiplexed mutation of GFP (x axis) and dsRed (y axis) in Hist1h3a -GFP Rosa26 -dsRed mouse ESCs (left) after co-introduction of Cas9, sgPal1 plasmid, and sgGFP and sgDsRed long-armed homology fragments (right) is shown. See also Figure S1 .

    Article Snippet: We vacuum centrifuged the DNA mixture to a final volume of < 20 μl and added 100 μl electroporation buffer from the Amaxa Human Stem Cell Nucleofector kit 1 to the human ESCs.

    Techniques: Flow Cytometry, Fluorescence, Electroporation, Plasmid Preparation, Microscopy, Knock-In, Mutagenesis

    Similar signaling capacity and cytotocixity of mRNA electroporated and retrovirally transduced T cells . ( a ) Sensitivity of T-cell activation using T cells produced by mRNA electroporation compared with retroviral transduction. Results are displayed as percentage of maximum IFN-γ response obtained from intracellular cytokine staining. ( b ) Dose dependent lysis of HepG2-env targets (solid symbols and line) or HepG2-core targets (open symbols and dotted line) by electroporated T cells (triangle) compared with retrovirally transduced T cells (circle). Results are displayed as mean of triplicate measurements + SD. ( c ) Expression of perforin (left panel) and granzyme (right panel) on a representative activated T cells electroporated (red histograms) or retrovirally transduced (blue histograms) with s183-TCR, after 5 hours coculture with peptide-loaded T2 cells. Coculture with unpulsed T2 cells served as negative control (gray histograms). MFI of perforin and granzyme are indicated. ( d ) mRNA electroporated T cells have T CM -like phenotype. Phenotype of total lymphocytes (left panel), CD8 + pentamer + T cells (middle panel) and CD4 + Vb3 + T cells (right panel) in s183-TCR mRNA electroporated T cells (red shaded area) compared with retrovirally transduced T cells (blue shaded area). The percentages of CD45RA +/- and CD62L +/- cells within total lymphocytes or the gated CD8 + pentamer + or CD4 + Vb3 + populations were determined by FACS. Cells were classified into different subsets: naive (CD45RA + CD62L + ), T CM (CD45RA − CD62L + ), T EM (CD45RA − CD62L − ) and terminally differentiated EM (CD45RA + CD62L − ). Results expressed as mean + SD ( n = 3).

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: A Practical Approach to Immunotherapy of Hepatocellular Carcinoma Using T Cells Redirected Against Hepatitis B Virus

    doi: 10.1038/mtna.2013.43

    Figure Lengend Snippet: Similar signaling capacity and cytotocixity of mRNA electroporated and retrovirally transduced T cells . ( a ) Sensitivity of T-cell activation using T cells produced by mRNA electroporation compared with retroviral transduction. Results are displayed as percentage of maximum IFN-γ response obtained from intracellular cytokine staining. ( b ) Dose dependent lysis of HepG2-env targets (solid symbols and line) or HepG2-core targets (open symbols and dotted line) by electroporated T cells (triangle) compared with retrovirally transduced T cells (circle). Results are displayed as mean of triplicate measurements + SD. ( c ) Expression of perforin (left panel) and granzyme (right panel) on a representative activated T cells electroporated (red histograms) or retrovirally transduced (blue histograms) with s183-TCR, after 5 hours coculture with peptide-loaded T2 cells. Coculture with unpulsed T2 cells served as negative control (gray histograms). MFI of perforin and granzyme are indicated. ( d ) mRNA electroporated T cells have T CM -like phenotype. Phenotype of total lymphocytes (left panel), CD8 + pentamer + T cells (middle panel) and CD4 + Vb3 + T cells (right panel) in s183-TCR mRNA electroporated T cells (red shaded area) compared with retrovirally transduced T cells (blue shaded area). The percentages of CD45RA +/- and CD62L +/- cells within total lymphocytes or the gated CD8 + pentamer + or CD4 + Vb3 + populations were determined by FACS. Cells were classified into different subsets: naive (CD45RA + CD62L + ), T CM (CD45RA − CD62L + ), T EM (CD45RA − CD62L − ) and terminally differentiated EM (CD45RA + CD62L − ). Results expressed as mean + SD ( n = 3).

    Article Snippet: 2.0 × 108 activated T cells were suspended in 5 ml electroporation buffer (MaxCyte, Gaithersburg, MD); s183-TCR mRNA was added at 200 μg/ml.

    Techniques: Activation Assay, Produced, Electroporation, Transduction, Staining, Lysis, Expressing, Negative Control, FACS

    High level of TCR expression and multifunctionality of mRNA electroporated T cells produced in large-scale, clinical-grade conditions . A schematic illustrating cell numbers, efficiency, yield, and functionality of laboratory-grade (top row) vs. clinical-grade (bottom row) electroporation of T cells. Dot plot of CD8 and HLA-A2-HBs183-191 pentamer staining in s183-TCR mRNA electroporated T cells at 24 hours postelectroporation. Bar charts show the frequency of IFN-γ, TNF-α, and IL-2 producing cells out of CD8 or CD4 electroporated T cells.

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: A Practical Approach to Immunotherapy of Hepatocellular Carcinoma Using T Cells Redirected Against Hepatitis B Virus

    doi: 10.1038/mtna.2013.43

    Figure Lengend Snippet: High level of TCR expression and multifunctionality of mRNA electroporated T cells produced in large-scale, clinical-grade conditions . A schematic illustrating cell numbers, efficiency, yield, and functionality of laboratory-grade (top row) vs. clinical-grade (bottom row) electroporation of T cells. Dot plot of CD8 and HLA-A2-HBs183-191 pentamer staining in s183-TCR mRNA electroporated T cells at 24 hours postelectroporation. Bar charts show the frequency of IFN-γ, TNF-α, and IL-2 producing cells out of CD8 or CD4 electroporated T cells.

    Article Snippet: 2.0 × 108 activated T cells were suspended in 5 ml electroporation buffer (MaxCyte, Gaithersburg, MD); s183-TCR mRNA was added at 200 μg/ml.

    Techniques: Expressing, Produced, Electroporation, Staining

    The in vitro assessment IRE outcome on a non-cancerous cancer cell line (human dermal fibroblasts, HDFs) to irreversible electroporation. Viabilities were assessed at a range of electric field (500 to 1250 V/cm) and different number of pulses (10, 50 and 99). All data points are the result of n = 4 trials and are reported as the average ± standard deviation.

    Journal: Annals of biomedical engineering

    Article Title: Physical and chemical enhancement of and adaptive resistance to irreversible electroporation of pancreatic cancer

    doi: 10.1007/s10439-017-1932-3

    Figure Lengend Snippet: The in vitro assessment IRE outcome on a non-cancerous cancer cell line (human dermal fibroblasts, HDFs) to irreversible electroporation. Viabilities were assessed at a range of electric field (500 to 1250 V/cm) and different number of pulses (10, 50 and 99). All data points are the result of n = 4 trials and are reported as the average ± standard deviation.

    Article Snippet: For each IRE test, 400 µL of the prepared cell suspension was pipetted into an electroporation cuvette (FB102, Fisher Scientific) between the two plate electrodes (2 mm apart).

    Techniques: In Vitro, Electroporation, Standard Deviation

    In vitro assessment of (a) physical, (b and c) chemical enhancement and (d) adaptive resistance to irreversible electroporation of pancreatic cancer cells (AsPC-1). (a) Comparison of IRE treatments of AsPC-1 cells in suspension after pulse sequencing (baseline, pulse timing, or low frequency). (b) Comparison of IRE outcome of AsPC-1 cells in PBS, DMEM and RPMI 1640. (c) Glucose concentration is positively correlated with post-IRE viability. (d) Adaptive resistance of cells shown by comparison of post-IRE cellular viability between originally untreated and IRE-treated cells. All data points are the result reported as the average ± standard deviation. Statistical significance is indicated with asterisks: *P

    Journal: Annals of biomedical engineering

    Article Title: Physical and chemical enhancement of and adaptive resistance to irreversible electroporation of pancreatic cancer

    doi: 10.1007/s10439-017-1932-3

    Figure Lengend Snippet: In vitro assessment of (a) physical, (b and c) chemical enhancement and (d) adaptive resistance to irreversible electroporation of pancreatic cancer cells (AsPC-1). (a) Comparison of IRE treatments of AsPC-1 cells in suspension after pulse sequencing (baseline, pulse timing, or low frequency). (b) Comparison of IRE outcome of AsPC-1 cells in PBS, DMEM and RPMI 1640. (c) Glucose concentration is positively correlated with post-IRE viability. (d) Adaptive resistance of cells shown by comparison of post-IRE cellular viability between originally untreated and IRE-treated cells. All data points are the result reported as the average ± standard deviation. Statistical significance is indicated with asterisks: *P

    Article Snippet: For each IRE test, 400 µL of the prepared cell suspension was pipetted into an electroporation cuvette (FB102, Fisher Scientific) between the two plate electrodes (2 mm apart).

    Techniques: In Vitro, Electroporation, Sequencing, Concentration Assay, Standard Deviation

    Suppression of S. mansoni ABC transporter expression enhances responsiveness of adult worms to PZQ. Adult schistosomes were electroporated with siRNAs targeting luciferase (Control) or the S. mansoni ABC transporters SMDR2, SmMRP1, ABCA4, ABCB6, and MRP7/ABCC10 (KD), as described in Materials and Methods . Following electroporation, worms were incubated in schistosome medium for 2 days, and then sorted into 2–3 worm pairs per well in a 12-well plate. They were then incubated in medium alone plus 0.5% DMSO, or in medium plus 800 nM PZQ, as described in Fig. 1 , and subsequently analyzed for motility. n = 6 for the experiments without PZQ and n = 18 (KD) or 19 (Control) for the experiments in PZQ. Motility was measured as in Fig. 1 , and results for individual worms are plotted. ** indicates P

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Inhibition or Knockdown of ABC Transporters Enhances Susceptibility of Adult and Juvenile Schistosomes to Praziquantel

    doi: 10.1371/journal.pntd.0003265

    Figure Lengend Snippet: Suppression of S. mansoni ABC transporter expression enhances responsiveness of adult worms to PZQ. Adult schistosomes were electroporated with siRNAs targeting luciferase (Control) or the S. mansoni ABC transporters SMDR2, SmMRP1, ABCA4, ABCB6, and MRP7/ABCC10 (KD), as described in Materials and Methods . Following electroporation, worms were incubated in schistosome medium for 2 days, and then sorted into 2–3 worm pairs per well in a 12-well plate. They were then incubated in medium alone plus 0.5% DMSO, or in medium plus 800 nM PZQ, as described in Fig. 1 , and subsequently analyzed for motility. n = 6 for the experiments without PZQ and n = 18 (KD) or 19 (Control) for the experiments in PZQ. Motility was measured as in Fig. 1 , and results for individual worms are plotted. ** indicates P

    Article Snippet: Briefly, following an overnight incubation in schistosome medium, adult worms (5 males plus 5 females) were placed in a 0.4 cm electroporation cuvette (USA Scientific, Ocala, FL) containing 50 µl siPORT (Life Technologies, Grand Island, NY) plus 3 µg of each of the siRNAs (IDT, Coralville, IA), either singly or in combination, targeting SMDR2, SmMRP1, MRP7, ABCA4, and ABCB6, or up to 15 µg luciferase siRNA (Life Technologies, Grand Island, NY; 3 µg per experimental siRNA used).

    Techniques: Expressing, Luciferase, Electroporation, Incubation

    Electroporation of vertebrate and mosquito cell lines with EILV reporter chimeras. RNA was transcribed in vitro , each cell line was electroporated with ∼10 μg of RNA, and phase-contrast (left) and fluorescence (right) micrographs were taken at 1 and 4 days postelectroporation (dpe).

    Journal: Journal of Virology

    Article Title: Novel Insect-Specific Eilat Virus-Based Chimeric Vaccine Candidates Provide Durable, Mono- and Multivalent, Single-Dose Protection against Lethal Alphavirus Challenge

    doi: 10.1128/JVI.01274-17

    Figure Lengend Snippet: Electroporation of vertebrate and mosquito cell lines with EILV reporter chimeras. RNA was transcribed in vitro , each cell line was electroporated with ∼10 μg of RNA, and phase-contrast (left) and fluorescence (right) micrographs were taken at 1 and 4 days postelectroporation (dpe).

    Article Snippet: Cells were mixed with each transcription mixture (∼10 μg of RNA), placed in 2-mm or 4-mm (Vero cells only) electroporation cuvettes, and immediately electroporated (BTX ECM-830 Square Wave electroporation system; Harvard Apparatus Inc., Holliston, MA) using the following conditions: 680 V, pulse length of 99 μs, interval between pulses of 200 ms, and number of pulses of 5.

    Techniques: Electroporation, In Vitro, Fluorescence

    Overexpression of Ascl1 changes the migratory behavior of neurons in the dorsal telencephalon. Ascl1 or US2 control expression construct were co-electroporated with a GFP expression plasmid. Two days after electroporation to the dorsal telencephalon, brains of E17.5 rats were dissected and sectioned in the coronal plane for slice culture and live imaging recording. The video from 5 to 11 hours after recording was used for tracking the migratory behavior. We set 0° to 180° axis in parallel to the lateral ventricle and the dorsomedial side as 180°. The line connecting the cell body location in the first frame (starting point) and the last frame (end point) of the video was used for measuring migratory angle. The average migratory rate was calculated as accumulated distance of every six-minute interval divided by recording time. 50 GFP-positive cells were counted for the control and 32 were counted for Ascl1 group. All counts were plotted into histograms according to their migratory angle or rate. ( A ) In the control group, most GFP-positive cells migrated radially in a migratory angle between 90° to 130°. ( B ) In Ascl1 group, GFP-positive cells were categorized into radially (90°–130°) and tangentially (140°–180°) migrating cells. ( C ) In the control group, GFP-positive cells migrated in the rate of 12.9 ± 3.6 μm/hour (mean ± SEM). ( D ) In Ascl1 group, radially migrating cells migrated in the rate of 30.2 ± 8.7 μm/hour and tangentially migrating cells migrated in the rate of 40.8 ± 9.3 μm/hour.

    Journal: Scientific Reports

    Article Title: Ascl1 promotes tangential migration and confines migratory routes by induction of Ephb2 in the telencephalon

    doi: 10.1038/srep42895

    Figure Lengend Snippet: Overexpression of Ascl1 changes the migratory behavior of neurons in the dorsal telencephalon. Ascl1 or US2 control expression construct were co-electroporated with a GFP expression plasmid. Two days after electroporation to the dorsal telencephalon, brains of E17.5 rats were dissected and sectioned in the coronal plane for slice culture and live imaging recording. The video from 5 to 11 hours after recording was used for tracking the migratory behavior. We set 0° to 180° axis in parallel to the lateral ventricle and the dorsomedial side as 180°. The line connecting the cell body location in the first frame (starting point) and the last frame (end point) of the video was used for measuring migratory angle. The average migratory rate was calculated as accumulated distance of every six-minute interval divided by recording time. 50 GFP-positive cells were counted for the control and 32 were counted for Ascl1 group. All counts were plotted into histograms according to their migratory angle or rate. ( A ) In the control group, most GFP-positive cells migrated radially in a migratory angle between 90° to 130°. ( B ) In Ascl1 group, GFP-positive cells were categorized into radially (90°–130°) and tangentially (140°–180°) migrating cells. ( C ) In the control group, GFP-positive cells migrated in the rate of 12.9 ± 3.6 μm/hour (mean ± SEM). ( D ) In Ascl1 group, radially migrating cells migrated in the rate of 30.2 ± 8.7 μm/hour and tangentially migrating cells migrated in the rate of 40.8 ± 9.3 μm/hour.

    Article Snippet: Forceps electrodes (7 mm in diameter for rats and 5 mm for mice; Harvard Apparatus) were used and five electric pulses separated by 500 ms were transmitted at 50 V for rats and 40 V for mice by an electroporation generator (Harvard Apparatus).

    Techniques: Over Expression, Expressing, Construct, Plasmid Preparation, Electroporation, Imaging

    Ephrin-A5 repulses cortical interneurons. Control (US2) or EfnA5 expression constructs were electroporated into the dorsal telencephalon together with a DsRed expression construct at E14.5 of Gad67-GFP mice. Brains were dissected four days after electroporation and sectioned coronally. ( A , B ) Confocal image of E18.5 mouse telencephalon. The cortex was divided into three regions: (A”,B”) the “in” region that contained DsRed-positive cells; (A”’,B”’) the “pre” region was ventrolateral to the “in” region; (A’,B’) the “post” region was dorsomedial to the “in” region. ( A , B ) In Ephrin-A5 group, fewer GFP-positive cells were present in “post” regions than those of the control group. Length of the scale bar is 100 μm. ( C ) Quantification of GFP fluorescent intensity in the MZ. ( D ) Quantification of GFP-positive cells in the CP, IZ and VZ/SVZ. A 200 μm wide cortical area in each region was selected for quantification. For the pre-DsRed, we selected the 200 μm wide cortical area 50 to 100 μm from the pre-/in-DsRed boundary. For the in-DsRed, we selected 50 to 100 μm from the in-/post-DsRed boundary. For the post-DsRed, we selected 50 to 100 μm from the in-/post-DsRed boundary. Data are presented as box and whisker plots and analyzed by using Student’s t-test, n = 4. * p

    Journal: Scientific Reports

    Article Title: Ascl1 promotes tangential migration and confines migratory routes by induction of Ephb2 in the telencephalon

    doi: 10.1038/srep42895

    Figure Lengend Snippet: Ephrin-A5 repulses cortical interneurons. Control (US2) or EfnA5 expression constructs were electroporated into the dorsal telencephalon together with a DsRed expression construct at E14.5 of Gad67-GFP mice. Brains were dissected four days after electroporation and sectioned coronally. ( A , B ) Confocal image of E18.5 mouse telencephalon. The cortex was divided into three regions: (A”,B”) the “in” region that contained DsRed-positive cells; (A”’,B”’) the “pre” region was ventrolateral to the “in” region; (A’,B’) the “post” region was dorsomedial to the “in” region. ( A , B ) In Ephrin-A5 group, fewer GFP-positive cells were present in “post” regions than those of the control group. Length of the scale bar is 100 μm. ( C ) Quantification of GFP fluorescent intensity in the MZ. ( D ) Quantification of GFP-positive cells in the CP, IZ and VZ/SVZ. A 200 μm wide cortical area in each region was selected for quantification. For the pre-DsRed, we selected the 200 μm wide cortical area 50 to 100 μm from the pre-/in-DsRed boundary. For the in-DsRed, we selected 50 to 100 μm from the in-/post-DsRed boundary. For the post-DsRed, we selected 50 to 100 μm from the in-/post-DsRed boundary. Data are presented as box and whisker plots and analyzed by using Student’s t-test, n = 4. * p

    Article Snippet: Forceps electrodes (7 mm in diameter for rats and 5 mm for mice; Harvard Apparatus) were used and five electric pulses separated by 500 ms were transmitted at 50 V for rats and 40 V for mice by an electroporation generator (Harvard Apparatus).

    Techniques: Expressing, Construct, Mouse Assay, Electroporation, Whisker Assay

    Overexpression of Ascl1 in the dorsal telencephalon induces ectopically tangential migration. Four days after electroporation to the dorsal telencephalon, brains of E19.5 rats were dissected and sectioned in the coronal plane. Electroporated cells were labeled with anti-GFP in green; nuclear DNA was stained with DAPI in blue. ( A ) A confocal image with a red square indicating the electroporated site. ( B’–G’ ) are zoomed regions of ( B–G ) indicated by red squares. ( B ) In the control group. GFP-positive cells were distributed near the electroporated area and many of them extended processes radially. GFP-positive axons toward the contralateral side were observed (yellow dashed lines). ( C ) In Neurog2 group, most GFP-positive cells were distributed in the cortical plate (CP). GFP-positive axons toward the contralateral side were observed (yellow dashed lines). ( D ) In Ascl1 group, many GFP-positive cells were distributed in the VZ/SVZ (white arrows) and IZ (white arrowheads) dorsomedially to the electroporated site. ( E ) In Dlx2 group, many GFP-positive cells were distributed in the VZ/SVZ (white arrows) dorsomedially to the electroporated site. ( F ) In Ascl1 + shLacZ#1 group, GFP-positive cells were distributed in a similar pattern as Ascl1 group in ( D ). ( G ) In Ascl1+shDlx2#1 group, many GFP-positive cells were distributed in the IZ (white arrowheads) dorsomedially to the electroporated site. Few GFP-positive cells were distributed in the VZ/SVZ (white arrows). Length of the scale bar is 120 μm in ( B – G ), and 100 μm in ( B’ to G’ ). ( H ) GFP-positive cells were categorized into radial, tangential, or other types according to the orientation of their leading processes. ( I ) Quantification of GFP-positive cells in the VZ/SVZ and IZ. Data are presented as box and whisker plots with all data points. The horizontal line within the box indicates the median, boundaries of the box indicate the 25th and 75th percentiles, and the whiskers indicate the maximum and minimum values of the results. Data are analyzed by using Student’s t-test, n = 6 in all groups. * p

    Journal: Scientific Reports

    Article Title: Ascl1 promotes tangential migration and confines migratory routes by induction of Ephb2 in the telencephalon

    doi: 10.1038/srep42895

    Figure Lengend Snippet: Overexpression of Ascl1 in the dorsal telencephalon induces ectopically tangential migration. Four days after electroporation to the dorsal telencephalon, brains of E19.5 rats were dissected and sectioned in the coronal plane. Electroporated cells were labeled with anti-GFP in green; nuclear DNA was stained with DAPI in blue. ( A ) A confocal image with a red square indicating the electroporated site. ( B’–G’ ) are zoomed regions of ( B–G ) indicated by red squares. ( B ) In the control group. GFP-positive cells were distributed near the electroporated area and many of them extended processes radially. GFP-positive axons toward the contralateral side were observed (yellow dashed lines). ( C ) In Neurog2 group, most GFP-positive cells were distributed in the cortical plate (CP). GFP-positive axons toward the contralateral side were observed (yellow dashed lines). ( D ) In Ascl1 group, many GFP-positive cells were distributed in the VZ/SVZ (white arrows) and IZ (white arrowheads) dorsomedially to the electroporated site. ( E ) In Dlx2 group, many GFP-positive cells were distributed in the VZ/SVZ (white arrows) dorsomedially to the electroporated site. ( F ) In Ascl1 + shLacZ#1 group, GFP-positive cells were distributed in a similar pattern as Ascl1 group in ( D ). ( G ) In Ascl1+shDlx2#1 group, many GFP-positive cells were distributed in the IZ (white arrowheads) dorsomedially to the electroporated site. Few GFP-positive cells were distributed in the VZ/SVZ (white arrows). Length of the scale bar is 120 μm in ( B – G ), and 100 μm in ( B’ to G’ ). ( H ) GFP-positive cells were categorized into radial, tangential, or other types according to the orientation of their leading processes. ( I ) Quantification of GFP-positive cells in the VZ/SVZ and IZ. Data are presented as box and whisker plots with all data points. The horizontal line within the box indicates the median, boundaries of the box indicate the 25th and 75th percentiles, and the whiskers indicate the maximum and minimum values of the results. Data are analyzed by using Student’s t-test, n = 6 in all groups. * p

    Article Snippet: Forceps electrodes (7 mm in diameter for rats and 5 mm for mice; Harvard Apparatus) were used and five electric pulses separated by 500 ms were transmitted at 50 V for rats and 40 V for mice by an electroporation generator (Harvard Apparatus).

    Techniques: Over Expression, Migration, Electroporation, Labeling, Staining, Whisker Assay

    Knockdown of Ephb1 or Ephb2 disrupted tangential migration promoted by Ascl1. Four days after electroporation to the dorsal telencephalon, brains of E19.5 rats were dissected and sectioned in the coronal plane. Electroporated cells were labeled with anti-GFP in green. ( A’–C’ ) are zoomed regions of ( A – C ) indicated by red squares, which were dorsomedial to the electroporated site. The cortex was divided equally into 10 bins. ( A ) In Ascl1 + shLacZ#1 group, many GFP positive cells were distributed in the VZ/SVZ (bins 1 and 2) and IZ (bin 5). ( B , C ) In Ascl1 + shEphB1#1 and Ascl1 + shEphB2#1 groups, GFP-positive cells were distributed in the VZ/SVZ (bins 1 and 2) and IZ (bin 5) were reduced. Length of the scale bar is 120 μm in ( A – C ), and 40 μm in ( A’–C’ ). ( D ) Distribution of GFP-positive in the cortex. Only GFP-positive cells dorsomedial to the electroporated site were counted. Data were presented as mean ± SEM with all data points and analyzed by one-way ANOVA with Tukey’s-HSD post hoc test, n = 4. * p

    Journal: Scientific Reports

    Article Title: Ascl1 promotes tangential migration and confines migratory routes by induction of Ephb2 in the telencephalon

    doi: 10.1038/srep42895

    Figure Lengend Snippet: Knockdown of Ephb1 or Ephb2 disrupted tangential migration promoted by Ascl1. Four days after electroporation to the dorsal telencephalon, brains of E19.5 rats were dissected and sectioned in the coronal plane. Electroporated cells were labeled with anti-GFP in green. ( A’–C’ ) are zoomed regions of ( A – C ) indicated by red squares, which were dorsomedial to the electroporated site. The cortex was divided equally into 10 bins. ( A ) In Ascl1 + shLacZ#1 group, many GFP positive cells were distributed in the VZ/SVZ (bins 1 and 2) and IZ (bin 5). ( B , C ) In Ascl1 + shEphB1#1 and Ascl1 + shEphB2#1 groups, GFP-positive cells were distributed in the VZ/SVZ (bins 1 and 2) and IZ (bin 5) were reduced. Length of the scale bar is 120 μm in ( A – C ), and 40 μm in ( A’–C’ ). ( D ) Distribution of GFP-positive in the cortex. Only GFP-positive cells dorsomedial to the electroporated site were counted. Data were presented as mean ± SEM with all data points and analyzed by one-way ANOVA with Tukey’s-HSD post hoc test, n = 4. * p

    Article Snippet: Forceps electrodes (7 mm in diameter for rats and 5 mm for mice; Harvard Apparatus) were used and five electric pulses separated by 500 ms were transmitted at 50 V for rats and 40 V for mice by an electroporation generator (Harvard Apparatus).

    Techniques: Migration, Electroporation, Labeling

    Overexpression of Ascl1 and Dlx2 in the ventral telencephalon promotes tangential migration. Four days after electroporation, brains of E19.5 rats were dissected and sectioned in the coronal plane. Electroporated cells were labeled with anti-GFP in green; nuclear DNA was stained with DAPI in blue. Red dashed lines indicate the electroporation region in ( A – G ). ( A ) A confocal image of brain section; red dashed lines mark the electroporated region and a red square indicate the area for ( B – D ). ST: striatum, LV: lateral ventricle. ( B – D ) GFP-positive cells were distributed in the ST adjacent to the LV. ( E ) In the control group, some GFP-positive cells were distributed in the VZ/SVZ (white arrows) of the dorsal telencephalon, while most remained in the ventral telencephalon. ( F ) In Ascl1 group, GFP-positive cells were distributed in both the VZ/SVZ (white arrows) and IZ (white arrowheads) of the dorsal telencephalon. ( G ) In Dlx2 group, many GFP-positive cells were distributed in the VZ/SVZ (white arrow). Red squares indicate the zoom-in areas for 1E’ to G’. Length of the scale bar is 40 μm in ( B – D ) and ( E’–G’ ), 250 μm in ( E – G ). ( H ) Quantification of GFP-positive cell density in the dorsal telencephalon. ( I ) Quantification of GFP-positive cells in the VZ/SVZ, IZ, or non-tangentially migrating cells in the dorsal telencephalon. Data were presented as mean ± standard error of the mean (SEM) with all data points and analyzed by Student’s t-test, n = 3. * p

    Journal: Scientific Reports

    Article Title: Ascl1 promotes tangential migration and confines migratory routes by induction of Ephb2 in the telencephalon

    doi: 10.1038/srep42895

    Figure Lengend Snippet: Overexpression of Ascl1 and Dlx2 in the ventral telencephalon promotes tangential migration. Four days after electroporation, brains of E19.5 rats were dissected and sectioned in the coronal plane. Electroporated cells were labeled with anti-GFP in green; nuclear DNA was stained with DAPI in blue. Red dashed lines indicate the electroporation region in ( A – G ). ( A ) A confocal image of brain section; red dashed lines mark the electroporated region and a red square indicate the area for ( B – D ). ST: striatum, LV: lateral ventricle. ( B – D ) GFP-positive cells were distributed in the ST adjacent to the LV. ( E ) In the control group, some GFP-positive cells were distributed in the VZ/SVZ (white arrows) of the dorsal telencephalon, while most remained in the ventral telencephalon. ( F ) In Ascl1 group, GFP-positive cells were distributed in both the VZ/SVZ (white arrows) and IZ (white arrowheads) of the dorsal telencephalon. ( G ) In Dlx2 group, many GFP-positive cells were distributed in the VZ/SVZ (white arrow). Red squares indicate the zoom-in areas for 1E’ to G’. Length of the scale bar is 40 μm in ( B – D ) and ( E’–G’ ), 250 μm in ( E – G ). ( H ) Quantification of GFP-positive cell density in the dorsal telencephalon. ( I ) Quantification of GFP-positive cells in the VZ/SVZ, IZ, or non-tangentially migrating cells in the dorsal telencephalon. Data were presented as mean ± standard error of the mean (SEM) with all data points and analyzed by Student’s t-test, n = 3. * p

    Article Snippet: Forceps electrodes (7 mm in diameter for rats and 5 mm for mice; Harvard Apparatus) were used and five electric pulses separated by 500 ms were transmitted at 50 V for rats and 40 V for mice by an electroporation generator (Harvard Apparatus).

    Techniques: Over Expression, Migration, Electroporation, Labeling, Staining

    Ephrin-A5 has a repulsive effect on Ascl1 -expressing cortical neurons. Control (US2) or Ascl1 expression constructs were electroporated into the dorsal telencephalon of E15.5 rats. The dorsal telencephalon was dissected two days after electroporation and dissociated into individual cells. These cells were cultured on coverslips coated with Fc-control or Ephrin-A5-Fc stripes. Cells were fixed 16–18 hours after plating and cells on strips or between strips were counted. Electroporated cells were labeled with anti-GFP in green, nuclear DNA was stained with DAPI in blue. GFP-positive cells on stripes are indicated by white arrowheads; GFP-positive cells between stripes are indicated by white arrows. ( A ) On a coverslip coated with Fc-control, GFP-positive Ascl1 -expressing cells and GFP-negative cells were distributed evenly on stripes and between stripes. ( B ) On a coverslip coated with Ephrin-A5-Fc, GFP-positive Ascl1 -expressing cells were preferentially distributed between stripes, while GFP-negative cells were evenly distributed on stripes and between stripes. Length of the scale bar is 50 μm. ( C , D ) Distribution of GFP-positive and GFP-negative cells. 150 cells from each group were counted in each experiment. Data are presented as mean ± SEM with all data points and analyzed by using Chi-square test, n = 3. * p

    Journal: Scientific Reports

    Article Title: Ascl1 promotes tangential migration and confines migratory routes by induction of Ephb2 in the telencephalon

    doi: 10.1038/srep42895

    Figure Lengend Snippet: Ephrin-A5 has a repulsive effect on Ascl1 -expressing cortical neurons. Control (US2) or Ascl1 expression constructs were electroporated into the dorsal telencephalon of E15.5 rats. The dorsal telencephalon was dissected two days after electroporation and dissociated into individual cells. These cells were cultured on coverslips coated with Fc-control or Ephrin-A5-Fc stripes. Cells were fixed 16–18 hours after plating and cells on strips or between strips were counted. Electroporated cells were labeled with anti-GFP in green, nuclear DNA was stained with DAPI in blue. GFP-positive cells on stripes are indicated by white arrowheads; GFP-positive cells between stripes are indicated by white arrows. ( A ) On a coverslip coated with Fc-control, GFP-positive Ascl1 -expressing cells and GFP-negative cells were distributed evenly on stripes and between stripes. ( B ) On a coverslip coated with Ephrin-A5-Fc, GFP-positive Ascl1 -expressing cells were preferentially distributed between stripes, while GFP-negative cells were evenly distributed on stripes and between stripes. Length of the scale bar is 50 μm. ( C , D ) Distribution of GFP-positive and GFP-negative cells. 150 cells from each group were counted in each experiment. Data are presented as mean ± SEM with all data points and analyzed by using Chi-square test, n = 3. * p

    Article Snippet: Forceps electrodes (7 mm in diameter for rats and 5 mm for mice; Harvard Apparatus) were used and five electric pulses separated by 500 ms were transmitted at 50 V for rats and 40 V for mice by an electroporation generator (Harvard Apparatus).

    Techniques: Expressing, Construct, Electroporation, Cell Culture, Labeling, Staining