Additional file 1: of Screening for potential genes associated with bone overgrowth after mid-shaft femur fracture in a rat model

All the DEGs screened between intact and fractured samples. (XLS 96 kb

Proteomic analysis of GBM cells by 2D-DIGE.

<p>A representative 2D-DIGE image (merged image) showed the protein profile of U251AR and U251 cells, labeled with Cy3 (green spots) and Cy5 (red spots), respectively. The approximate molecular weight range in the vertical dimension was from 10 to 150 kD. The PI of proteins ranged from 3 to 10. The differently expressed protein spot ID were indicated with white arrows. The protein spot 2421 and 1737 was PTRF and VIM, respectively.</p

The cell viability (IC50) changes after PTRF konckdown.

<p>(A) The cell viabilty of U251AR cells, U251AR cells transfected with shNC and U251AR cells transfected with shPTRF after treatment with 100 µg/mL TMZ for 24 h, 48 h, 72 h, 96 h, and 120 h; (B) The IC50 of U251AR cells and U251AR cells transfected with shNC and shPTRF; (C) The IC50 of U251 cells and U251 cells transfected with shNC and shPTRF. **, P<0.05.</p

The nucleotide sequences of the target miRNA and the unrelated miRNA.

<p>The nucleotide sequences of the target miRNA and the unrelated miRNA.</p

PTRF and caveolin1 expression in astrocytoma tissues as indicated by immunohistochemistry detection.

<p>(A) PTRF in normal tissues; (B) PTRF in grade I astrocytoma; (C) PTRF in grade II astrocytoma; (D) PTRF in grade III astrocytoma; (E) PTRF in primary GBM patients; (F) PTRF in relapsed GBM patients; (G) Caveolin1 in primary GBM patients; (H) Caveolin1 in relapsed GBM patients; (I) Negative control.</p

Establishment of the stablely transfected cells and the expression of PTRF in stable transfected cells.

<p>(A) Morphology of U251AR cells transfected with shPTRF (U251ARshPTRF cells); (B) Morphology of U251 cells transfected with shPTRF (U251shPTRF cells); Light microscopy, 20× (A, B); Fluorescence microscopy, 20× (A, B); (C, D) The protein and mRNA expression of PTRF, caveoin1, and P-gp in U251AR cells after transfection with shPTRF and negative vector (shNC) by Western bolt and quantitative RT-PCR. (E, F) The protein and mRNA expression of PTRF, caveoin1, and P-gp in U251 cells after transfection with shPTRF and shNC as indicated by Western blot analysis and quantitative RT-PCR.</p

Expression levels of PTRF and caveolin1 in GBM patients.

<p>(A) PTRF and Caveolin1 expression in primary GBM and relapsed GBM patients as detected by quantitative RT-PCR (*, P<0.05). (B) The correlation of PTRF and caveolin1 mRNA in the same GBM specimens (2-tailed Pearson correlation, r = 0.766, P<0.01).</p

Primer sequences used in quantitative RT-PCR analysis.

<p>Primer sequences used in quantitative RT-PCR analysis.</p

Twenty-one differentially expressed proteins in U251 cell line versus U251AR cell line.

<p>Note:</p>a<p>Protein MW, Protein molecular weight;</p>b<p>Fold changes (mean ± SD) of U251 cell line vs. U251AR cell line, which were calculated from the DeCyder/spot volume analysis;</p>c<p>up, up-regulated in the U251AR cell line;</p>d<p>down, down-regulated in the U251AR cell line.</p

The biological characteristics of MDR cell line U251AR.

<p>(A) The expression of P-gp was significantly increased in chemoresistant cell line U251AR as detected by Western blot analyses. (B) The mRNA levels of P-gp, MRP1, and BCRP in U251 and U251AR cells were determined by quantitative RT-PCR. Values of three independent experiments were represented as the mean ± SD. *, P<0.05.</p