Schematic diagram of the putative miR-9-3p binding site in the UTRs of Herpud1.

<p>The seed sequence of miR-9-3p matches the UTRs of Herpud1. The 3 mutated nucleotides of the Herpud1-UTR are in bold and underlined.</p

Herpud1 levels in U251 cells and glioma tissues.

<p>(A) U251 cells were transfected with 200 nM negative control miRNA (NC) and miR-9-3p mimics. Expression of Herpud1 was normalized by GAPDH using the 2^-ΔΔct method. **P<0.01; (B) and the protein level of HERPUD1 was detected by westernblot. (C) Relative expression levels of Herpud1 were measured by using qRT-PCR with GAPDH as an internal control. The box extends from the 25^th to the 75^th percentile; the bar in the middle indicates the 50^th percentile; the whisker caps indicate the maximum and the minimum values. S: significant; NS: not significant. *P<0.05.</p

MiR-9-3p augments H₂O₂ induced apoptosis through Herpud1.

<p>(A) Cell proliferation of transfected U251 cells was determined using CCK8 assay at 24h, 48 and 72h. U251 cells were treated with 500μM H₂O₂ for 4, 6, 8 and 24h after transfected with miR-9-3p mimics (B) or HERPUD1-RNAi plasmids (D) for 48h. Apoptotic cells were detected by staining with FITC-Annexin V and propidium iodide. (C) U251 cells were transfected with 2μg HERPUD1-RNAi-CON. and three RNAi plasmids. The protein level of HERPUD1 was detected by westernblot.</p

Heat map of differential miRNA expression in glioma and non-tumor tissues.

<p>Both down-regulated (red) and up-regulated (blue) miRNAs were identified in glioma tissues (N = 3) versus non-tumor tissues (N = 3).</p

MiR-9-3p augments apoptosis induced by H₂O₂ through down regulation of Herpud1 in glioma - Fig 5

<p>Relative luciferase activities of Herpud1 wild type (wt) UTRs (A) and three mutant UTRs of Herpud1 (B) were obtained by co-transfection of negative control miRNA or miR-9-3p mimics and pRL-CMV plasmid. Relative luciferase activity was calculated as the ratio of firefly/Renilla activities in the cells and normalized to those of the control. The results are presented as mean±SD from three independent experiments. ***P<0.001.</p

The primers for real-time PCR.

<p>The primers for real-time PCR.</p

Relative expression levels of miRNAs were measured in 24 glioma samples and 5 control samples by using quantitative RT-PCR with U6 as an internal control (normalized by U6 using 2^-Δct).

<p>The box extends from the 25^th to the 75^th percentile; the bar in the middle indicates the 50^th percentile; the whisker caps indicate the maximum and the minimum values. (A) miR-9-3p expression (P = 0.003); (B) miR-126 expression (P<0.0001); (C) miR-572 expression (P<0.0001).</p