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The fresh supernatant is got rid of (Profile 3E, lanes dos–5) and the beans had been blended with 20 µl away from 2× Laemmli sample buffer and incubated from the 90°C for five min and you may separated because of the SDS-Web page for the an excellent cuatro%–20% gradient gel. The fresh gel is sometimes gold stained (SilverQuest, Invitrogen) or moved to nitrocellulose for immunoblotting (understand the actions section inside Text message S1 to own details). The silver-tarnished proteins rings was slash and you may recognized by microcapillary drinking water chromatography combination size spectrometry (Taplin Bulk Spectrometry Studio, Harvard Scientific School). (1) mRNAs which includes ALREX-producing SSCRs is actually packaged on the mRNPs and you can exported because of the an unfamiliar procedure.
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