Journal: Journal of visualized experiments : JoVE

Article Title: Defining Gene Functions in Tumorigenesis by Ex Vivo Ablation of Floxed Alleles in Malignant Peripheral Nerve Sheath Tumor Cells

doi: 10.3791/62740

Figure Lengend Snippet: Table of Materials

Article Snippet: For the bioluminescence (MTT) assays, perform the assay exactly as outlined in the manufacturer’s instructions every 24 h for 72 h using a luminometer plate reader. table ft1 table-wrap mode="anchored" t5 caption a7 Name of Material/ Equipment Company Catalog Number Comments Ad5CMV-eGFP Gene Transfer Vector Core, Univ of Iowa VVC-U of Iowa-4 Ad5CMVCre-eGFP Gene Transfer Vector Core, Univ of Iowa VVC-U of Iowa-1174 alexa 568 secondary antibody Thermo/Fisher GαR A11036 Bioconductor Open Source Software for Bioninformatics Bioconductor http://www.bioconductor.org alternative statistical analysis tool used for step 4.4 CD31 Abcam ab28364 Celigo Image Cytometer Nexcelom Bioscience N/A Cell Stripper Corning 25-056-Cl mixture of chelators DAB staining kit Vector Labs MP-7800 DAVID ( D atabase for A nnotation, V isualization, and I ntegrated D iscovery) DAVID https://david.ncifcrf.gov functional enrichment analysis software used for step 4.5 DMEM Corning 15-013-Cl DreamTaq and Buffer (Genotyping PCR) Thermo/Fisher EP0701 and K1072 erbB4 antibodies Santa Cruz sc-284 erbB4 antibodies Abcam ab35374 erbB4 antibodies Millipore HFR1: 05-1133 FACS Sorter BD Biosciences Aria II Forskolin Sigma F6886 GenomeSpace Tools and Data Sources GenomeSpace https://genomespace.org/support/tools/ general resource for several types of open source bioinformatic tools for step 4.5 Glutamine Corning 25-005-Cl Gorilla G ene Ontology en RI chment ana L ysis and visua L iz A tion tool Gorilla N/A, http://cbl-gorilla.cs.technion.ac.il functional enrichment analysis software used for step 4.5 GSEA Gene Set Enrichment Analysis Broad Institute N/A, https://www.gsea-msigdb.org/gsea/index.jsp functional enrichment analysis software used for step 4.5 HSD: Athymic Nude-FOxn1nu mice Envigo (Previously Harlan Labs) 69 Illumina HiSeq2500 (next generation DNA sequencer) Illumina Hi Seq 2500 DNA sequencer used for step 4.2 Lasergene: ArrayStar Gene expression and variant analysis DNAStar LaserGene software N/A software statistical and normalization analysis used for step 4.4 Lasergene: SeqMan NGen sequence alignment assembly software alignment used for step 4.3 N/A software alignment used for step 4.3 Matrigel, low growth factor basement membrane matrix Corning 354230 NRG1-beta In house Generated by SLC, also commercially available from R & D Systems(396-HB-050/CF).

Techniques: Plasmid Preparation, Software, Cytometry, Staining, Functional Assay, Expressing, Variant Assay, Sequencing, Generated, Viability Assay, Amplification

Journal: Journal of visualized experiments : JoVE

Article Title: Defining Gene Functions in Tumorigenesis by Ex Vivo Ablation of Floxed Alleles in Malignant Peripheral Nerve Sheath Tumor Cells

doi: 10.3791/62740

Figure Lengend Snippet: Table of Materials

Article Snippet: For the bioluminescence (MTT) assays, perform the assay exactly as outlined in the manufacturer’s instructions every 24 h for 72 h using a luminometer plate reader. table ft1 table-wrap mode="anchored" t5 caption a7 Name of Material/ Equipment Company Catalog Number Comments Ad5CMV-eGFP Gene Transfer Vector Core, Univ of Iowa VVC-U of Iowa-4 Ad5CMVCre-eGFP Gene Transfer Vector Core, Univ of Iowa VVC-U of Iowa-1174 alexa 568 secondary antibody Thermo/Fisher GαR A11036 Bioconductor Open Source Software for Bioninformatics Bioconductor http://www.bioconductor.org alternative statistical analysis tool used for step 4.4 CD31 Abcam ab28364 Celigo Image Cytometer Nexcelom Bioscience N/A Cell Stripper Corning 25-056-Cl mixture of chelators DAB staining kit Vector Labs MP-7800 DAVID ( D atabase for A nnotation, V isualization, and I ntegrated D iscovery) DAVID https://david.ncifcrf.gov functional enrichment analysis software used for step 4.5 DMEM Corning 15-013-Cl DreamTaq and Buffer (Genotyping PCR) Thermo/Fisher EP0701 and K1072 erbB4 antibodies Santa Cruz sc-284 erbB4 antibodies Abcam ab35374 erbB4 antibodies Millipore HFR1: 05-1133 FACS Sorter BD Biosciences Aria II Forskolin Sigma F6886 GenomeSpace Tools and Data Sources GenomeSpace https://genomespace.org/support/tools/ general resource for several types of open source bioinformatic tools for step 4.5 Glutamine Corning 25-005-Cl Gorilla G ene Ontology en RI chment ana L ysis and visua L iz A tion tool Gorilla N/A, http://cbl-gorilla.cs.technion.ac.il functional enrichment analysis software used for step 4.5 GSEA Gene Set Enrichment Analysis Broad Institute N/A, https://www.gsea-msigdb.org/gsea/index.jsp functional enrichment analysis software used for step 4.5 HSD: Athymic Nude-FOxn1nu mice Envigo (Previously Harlan Labs) 69 Illumina HiSeq2500 (next generation DNA sequencer) Illumina Hi Seq 2500 DNA sequencer used for step 4.2 Lasergene: ArrayStar Gene expression and variant analysis DNAStar LaserGene software N/A software statistical and normalization analysis used for step 4.4 Lasergene: SeqMan NGen sequence alignment assembly software alignment used for step 4.3 N/A software alignment used for step 4.3 Matrigel, low growth factor basement membrane matrix Corning 354230 NRG1-beta In house Generated by SLC, also commercially available from R & D Systems(396-HB-050/CF).

Techniques: Plasmid Preparation, Software, Cytometry, Staining, Functional Assay, Gene Expression, Variant Assay, Sequencing, Membrane, Generated, Viability Assay, Amplification