Distinguishing parallel from series circuits with a double knockdown procedure in human cell lines.
Distinguishing parallel from series circuits with a double knockdown procedure in human cell lines.
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DOI:
10.1016/j.xpro.2022.101890
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发表时间:
2022-12-16
期刊:
影响因子:
--
通讯作者:
Edelman, Arthur M.
中科院分区:
文献类型:
--
作者:
Gocher-Demske, Angela M.;Dai, Shuhang;Edelman, Arthur M.
Signaling cascades can act in series or in parallel. Here, we describe a convenient and robust protocol for dual, sequential knockdown of two proteins using RNA interference. We detail the steps for a quantitative mapping of signaling circuitry. We used this approach to study kinases in human ovarian cancer cells, but the protocol can be applied to many other posttranslational modifications. For complete details on the use and execution of this protocol, please refer to Gocher et al. (2017). Knockdown of two proteins via a dual sequential RNA interference Quantitative distinction of in-parallel versus in-series signaling cascades Specific steps for kinases, but applicable to many posttranslational modifications Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Signaling cascades can act in series or in parallel. Here, we describe a convenient and robust protocol for dual, sequential knockdown of two proteins using RNA interference. We detail the steps for a quantitative mapping of signaling circuitry. We used this approach to study kinases in human ovarian cancer cells, but the protocol can be applied to many other posttranslational modifications.
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