Association of miR-124 to hStau1 complexes in undifferentiated and differentiated neuroblastoma cells.

Cultures of SH-SY5Y neuroblastoma cells were differentiated as described in Materials and Methods. Total cell extracts were isolated from cells prior to differentiation (day 0) or at a final stage of differentiation (day 10) and fractionated on a Sephacryl S-400 column. (A) The various fractions were analysed by Western-blot with antibodies specific for Ago2 and hStau1 The gels to analyse hStau1 were run longer than in Fig. 4 to better separate hStau1 55 and 63 kDa isoforms. (B) RNA was isolated from the fraction pools indicated and the concentration of miR-124 was determined by TaqMan RT-qPCR. (C) The amounts of hStau155 and hStau163 isoforms were determined by Western-blot (see A) and their ratios is presented as average and standard deviations of 3 determinations. * indicates a p-value<0.05 in a two-tailed Student's t-test.