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In 2016, as part of the Reproducibility Project: Cancer Biology, we published a Registered Report (Fiehn et al., 2016), that described how we intended to replicate selected experiments from the paper "The common feature of leukemia-associated IDH1 and IDH2 mutations is a neomorphic enzyme activity converting alpha-ketoglutarate to 2-hydroxyglutarate" (Ward et al., 2010). Here, we report the results of those experiments. We found that cells expressing R172K mutant IDH2 did not display isocitrate-dependent NADPH production above vector control levels, in contrast to the increased production observed with wild-type IDH2. Conversely, expression of R172K mutant IDH2 resulted in increased alpha-ketoglutarate-dependent consumption of NADPH compared to wild-type IDH2 or vector control. These results are similar to those reported in the original study (Figure 2; Ward et al., 2010). Further, expression of R172K mutant IDH2 resulted in increased 2HG levels within cells compared to the background levels observed in wild-type IDH2 and vector control, similar to the original study (Figure 3D; Ward et al., 2010). In primary human AML samples, the 2HG levels observed in samples with mutant IDH1 or IDH2 status were higher than those observed in samples without an IDH mutation, similar to what was observed in the original study (Figure 5C; Ward et al., 2010). Finally, we report meta-analyses for each result.
Pubmed ID: 28653623 RIS Download
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View all literature mentionsNon-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.
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View all literature mentionsThis unknown targets Immun-Star Goat Mouse (GAM)-HRP Conjugate
View all literature mentionsThis polyclonal secondary targets IgG (H+L)
View all literature mentionsThis monoclonal targets α-tubulin
View all literature mentionsThis monoclonal targets IDH2
View all literature mentionsThis polyclonal targets Idh1
View all literature mentionsSoftware that configures and controls the measurement protocols and actions performed by the FilterMax Multi-Mode Microplate Readers and the SpectraMax Paradigm Multi-Mode Detection Platform, which are commercially available microplate readers from Molecular Devices.
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View all literature mentionsA software used for quantitative Western blot imaging and analysis. This software is compatible with several dyes and kits, providing extra flexibility for application. This software is designed to be able to look at multiple proteins simultaneously on a single assay. This software is sensitive enough to distinguish co-migrating proteins, such as phosphorylated and unphosphorylated isoforms.
View all literature mentionsCell line HEK293T is a Transformed cell line with a species of origin Homo sapiens (Human)
View all literature mentionsPlatform to support research and enable collaboration. Used to discover projects, data, materials, and collaborators helpful to your own research.
View all literature mentionsSoftware that configures and controls the measurement protocols and actions performed by the FilterMax Multi-Mode Microplate Readers and the SpectraMax Paradigm Multi-Mode Detection Platform, which are commercially available microplate readers from Molecular Devices.
View all literature mentionsSoftware that configures and controls the measurement protocols and actions performed by the FilterMax Multi-Mode Microplate Readers and the SpectraMax Paradigm Multi-Mode Detection Platform, which are commercially available microplate readers from Molecular Devices.
View all literature mentionsSoftware environment and programming language for statistical computing and graphics. R is integrated suite of software facilities for data manipulation, calculation and graphical display. Can be extended via packages. Some packages are supplied with the R distribution and more are available through CRAN family.It compiles and runs on wide variety of UNIX platforms, Windows and MacOS.
View all literature mentionsA software used for quantitative Western blot imaging and analysis. This software is compatible with several dyes and kits, providing extra flexibility for application. This software is designed to be able to look at multiple proteins simultaneously on a single assay. This software is sensitive enough to distinguish co-migrating proteins, such as phosphorylated and unphosphorylated isoforms.
View all literature mentionsThis unknown targets Immun-Star Goat Mouse (GAM)-HRP Conjugate
View all literature mentionsThis polyclonal secondary targets IgG (H+L)
View all literature mentionsThis monoclonal targets α-tubulin
View all literature mentionsThis monoclonal targets IDH2
View all literature mentionsThis polyclonal targets Idh1
View all literature mentionsSoftware that configures and controls the measurement protocols and actions performed by the FilterMax Multi-Mode Microplate Readers and the SpectraMax Paradigm Multi-Mode Detection Platform, which are commercially available microplate readers from Molecular Devices.
View all literature mentionsCell line HEK293T is a Transformed cell line with a species of origin Homo sapiens (Human)
View all literature mentionsPlatform to support research and enable collaboration. Used to discover projects, data, materials, and collaborators helpful to your own research.
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