Fig. 1.

Rapamycin upregulates miR-29b expression in Tsc2-deficient but not Tsc2 wild-type cells in vitro and in vivo. miR-29b expression was assessed by RT-qPCR in Tsc2^+/+ or Tsc2^−/− MEFs (a), Wild-type or Tsc2 knockout MEFs (b), and in Tsc2-deficient rat ERL4 cells (c), treated with vehicle (DMSO) or rapamycin (20 nM) for 24 h. Data are presented as mean fold change in miR-29b expression ± SD relative to Tsc2^+/+ MEFs, Tsc2 WT MEFs, or ERL4 treated with vehicle. Results are from n = 3 biological replicates. Data are presented as mean ± SD. Data for bar graphs were calculated using two-way ANOVA followed by Bonferroni’s posttest for multiple comparisons. d Mice bearing ERL4 xenograft tumors were treated intraperitoneally with rapamycin (3 mg/kg) or vehicle every other day for 6 days (vehicle: n = 3, rapamycin: n = 4). Tumors were harvested 4 h after the last treatment and miR-29b expression was examined by RT-qPCR. Data are presented as mean ± SD. Statistical significance was determined by Mann–Whitney’s U test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001