Figure 2. miR-21 and miR-199a-3p target mTor and Pten mRNAs. A, B, RTddPCR for Pten (A) and mTor (B) mRNAs from precipitations of 3’biotinylated miR-21, miR-199a-3p or scrambled RNA (control) transfected DRG neurons is shown as average relative to total input mRNA ± SEM (N ≥ 4; **p ≤ 0.01 vs control and ##p ≤ 0.01 vs miR-21 by one-way ANOVA with pair-wise comparison and Tukey’s post hoc tests). C, D, Transfection of precursor miR-21 and miR-199a-3p (pre-miR-21 and pre-miR-199a-3p, respectively) into DRG neurons increased levels of mature miR-21 (C) and miR-199a-3p levels (D). E–G, DRG cultures transfected with pre-miR-21 showed decrease in Pten mRNA by RTddPCR (E) and PTEN protein by immunoblotting (F, G). F, Representative immunoblot for PTEN protein, with GAPDH immunoblot showing relatively equal loading between lanes. E, K, Average ± SEM. H, Anti-miR-21 transfection in adult DRG neuron cultures increases levels of Pten mRNA. I–K, DRG cultures transfected with pre-miR-199a-3p showed decrease in mTor mRNA by RTddPCR (I) and mTOR protein by immunoblotting (J, K). F, Representative immunoblot for mTOR protein, with GAPDH immunoblot showing relatively equal loading between lanes. I, K, Average values ± SEM. L, Anti-miR-199a-3p transfection in adult DRG neuron cultures increases levels of mTor mRNA (N = 4 for E, H–J; N = 3 for G, K; **p ≤ 0.01 and ***p ≤ 0.005, vs control by Student’s t test).