FIG. 10.

Trio expression is lowered in FLNc-deficient cells. (A) Quantitative RT-PCR analysis of Trio transcript levels in FLNc-deficient cells (F8) versus control cells throughout myogenic differentiation. On day 0, cells were switched to low-serum conditions. All amounts were normalized to an internal control (Tbp) and then to the amount of transcription in control cells at day 3. The data shown are the averages of results from three experiments, and error bars represent standard errors. (B) The amount of Trio knockdown was determined using quantitative RT-PCR. All of the amounts were first normalized to the endogenous Tbp (TATA binding protein) transcript levels and then to the control cell line. The cells were allowed to differentiate for 8 days. Experiments were performed in triplicate, and the error bars represent standard errors. The Trio2 line shows a decrease of 95% in Trio transcripts. (C) Ten micrograms of total protein isolated from the C2C12 cell lines was loaded onto 3 to 8% Tris-acetate gels (Invitrogen), and Western analysis was performed using anti-TRIO antibodies. Antibodies against β-actin were used to control for even loading. The Trio2 line has virtually no TRIO protein expressed.