FIG 6.
RasG12V inhibits TXNIP translation elongation independently of TXNIP's mRNA primary sequence. (A) Chromatograms showing the polysome profiles of MondoA KO::TXNIP::Vector (Vector) or RasG12V (Ras) MEFs before (−Harr) and after (+Harr) harringtonine treatment. Fraction numbers are shown on the x axis, with heavy-polysome fractions shaded in yellow. (B to E) RT-qPCR was used to determine the amounts of TXNIP-WT mRNA (B), TXNIP-MUT mRNA (C), TXNIP-OPT mRNA (D), and actin mRNA (E) from heavy-polysome fractions of the listed cell population. The amounts of heavy-polysome-associated mRNAs were normalized to that of total RNA. The percent decrease in heavy-polysome-associated RNA levels caused by harringtonine treatment was calculated for each cell line. Experiments were repeated twice, and a representative experiment is shown. Values are reported as means and SD. Statistical significance was determined using one-way ANOVA. **, P < 0.01; ***, P < 0.001; ns, P ≥ 0.05.