Figure 3

FMRP expression is maintained when cap-dependent translation is reduced by rapamycin or eIF-4E siRNA. (A) Lysates from untreated or rapamycin treated C6 cells were analyzed for FMRP, phosphorylated p70 S6 kinase, and eIF-4E. Shown is a representative Western blot for each experiment, n = 3. (B) siRNA directed against eIF-4E or a nonsense siRNA were transfected into C6 cells for 72 hr. Lysates from these cells were analyzed for eIF-4E, FMRP, JunB and GAPDH. GAPDH was used as a loading control since it has an extended half-life. Shown is a representative Western blot for each experiment, n = 3. (C) Western blots of lysates from cells treated with eIF-4E or a nonsense siRNA. Expression level of the proteins were quantitated using ImageQuant software (n = 3) and were normalized to GAPDH levels.