Detail
| Localization |
Tissue |
Validated Method |
PMID |
| Endoplasmic reticulum |
U2OS cell |
Fluorescence in situ hybridization |
26272916 |
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Description: Next, we monitored the localization of nesprin-2 (SYNE2) mRNA, which encodes a giant tail-anchored protein (796 kDa) that is present on the outer nuclear envelope and is involved in nuclear positioning (Luxton et al., 2010). After extraction, about two thirds of the foci remained, indicating that some of this mRNA was anchored to the ER (Fig. 1A,B). To ensure that the FISH signal was specific, we also probed cells that were depleted of the endogenous nesprin-2 mRNA using RNA interference (RNAi). Indeed, small hairpin RNA (shRNA)-treated cells lost 90% of their signal (supplementary material Fig. S1), indicating that our nesprin-2 probes detected the intended target. Like Sec61b, nesprin-2 mRNA largely remained associated with the ER in cells treated with HHT, or puromycin and EDTA. Thus nesprin-2, like Sec61b, can associate with the ER membrane, and this activity is mostly independent of translation.
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Other Subcellular Localization: