Abstract
Export of transcribed mRNAs from nucleoplasm to cytosol is an essential process for the translation of genes into proteins. This process is tightly regulated by nuclear pores, composed of about 30 nucleoporin proteins (Nups). Whether or not the mRNAs are able to be appropriately exported to cytoplasm is of an importance for understanding the role of Nups. Here, we describe a practical protocol to detect the intracellular localization of mRNAs in mesophyll cells of Nicotiana benthamiana (N. benthamiana). This protocol is based on poly (A) in situ hybridization method using an oligo d(T) probe conjugated with Alexa Fluor-488.
Keywords: poly (A) in situ hybridization, Nicotiana benthamiana, Virus-induced gene silencing, nucleoporin, mRNAs
Materials and Reagents
Equipment
Procedure
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Acknowledgments
This protocol was adapted from Parry et al. (2006) and Germain et al. (2010). The work was supported by a Grant-in-Aid for Scientific Research (B) (26292024) from the Japan Society for the Promotion of Science and by Grant for Basic Science Research Projects from the Sumitomo Foundation.
References
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