Figure 4.

miR-7 and miR-153 activated p70S6K signaling cascade in cortical neurons. Six to seven days old primary cortical neurons were transduced with adenoviral particles expressing scramble miR, miR-7, or miR-153 and were lysed 48 h post-transduction. (A) Equal amounts of total protein from lysates of cortical neurons were analyzed on 10% SDS-PAGE and immunoblotted with antibodies specific for phosphorylated forms of AKT, p38 MAPK, ERK1/2 SAPK/JNK. To ensure equal loading membranes were re-probed against AKT, p38 MAPK, ERK1/2, and SAPK/JNK, respectively. (B,C) Equal amounts of total protein from lysates of cortical neurons were analyzed on 10% SDS-PAGE and immunoblotted with antibodies specific for phosphorylated forms of p70S6K (B) as well as for the phosphorylated forms of p70S6K substrate, S6RP (C). To ensure equal loading, membranes were re-probed against p70S6K and S6RP, respectively. Quantification of the results was performed by scanning densitometry. Bars in the graph depict mean ± s.e.m. Note that compared to scramble-transduced controls, primary cortical neurons transduced with miR-7 or miR-153 expressing adenoviruses displayed a significant increase only in the levels of phosphorylated forms of p70S6K and S6RP. ^**P ≤ 0.01, ^***P ≤ 0.001.