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Analysis of hStau1 complexes and the associated miRNAs in human neuroblastoma cells.

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posted on 25.11.2014 by Joan Peredo, Patricia Villacé, Juan Ortín, Susana de Lucas

Soluble extracts derived from the SH-SY5Y neuroblastoma cell line were filtered on a Sephacryl S-400 column. (A) The various fractions were analysed by Western-blot with antibodies specific for Ago2, RCK/p54 and hStau1. (B) The fractions were grouped in 4 pools: F1 (17, 18), F2 (21,22,23), F3 (26,27,28,29) and F4 (34,35,36 as a negative control). RNA was isolated in each pool and analysed by TaqMan RT-qPCR to quantify the six miRNAs most prevalent in hStau1 complexes. (C) The amounts of miR-124 and miR-9 were determined in the fractions pools F1 to F4 derived from 3 independent filtration experiments. Values are averages and standard deviations and represent the amount of miRNA present in each fraction pool as percentage of total miRNA recovered from F1+F2+F3+F4 pools.