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. 2022 Sep 15;8(5):62. doi: 10.3390/ncrna8050062

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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iDRIP-MS reveals direct lnc-Nr6a1-1-interacting proteins. (A) A schematic overview of the iDRIP-MS method. (B) Retrieval of RNA by iDRIP using lnc-Nr6a1-1 and LacZ probes measured by RT-qPCR. Efficiency of lnc-Nr6a1-1 pulldown was determined by comparing to a standard curve generated using serial dilutions of input. Error bars S.D. (C) Selected lnc-Nr6a1-1-interacting proteins identified by iDRIP are grouped into functional classes. (D) Physical association of lnc-Nr6a1-1 with Lamin A/C, g-catenin (Plakoglobin), ENO1, ALDOA and GAPDH was confirmed by RNA immunoprecipitation (RIP). ENO1, enolase; ALDOA, aldolase; GAPDH, glyceraldehyde 3-phophatedeshydrogenase. The results are the median of three technical replicates; error bars represent S.D. (E) Subcellular localization of lnc-Nr6a1 isoforms. Expression of Hprt, lnc-Nr6a1-1, lnc-Nr6a1-2, U3 and Xist in cytoplasmic, nuclear soluble and nuclear insoluble fractions of NMuMG cells. The graph is an average of three technical replicates; error bars represent S.D.