Fig 1: RNA decoy oligonucleotides targeting YB-1 inhibited tumor cell growth in vivo.(A and B) MTT analysis of cell growth in U251 (A) and U87 (B) cells expressing control or YB-1–specific shRNA with or without control or YB-1–specific (YBX1-1) decoy oligonucleotides. Data are presented as mean ± SEM (n = 3). ***P < 0.001 by 1-way ANOVA followed by Dunnett’s test. (C) SDS-PAGE of material pulled down without added oligonucleotides (mock) or with biotinylated scrambled or YB-1–specific (YBX1-2) decoy oligonucleotides followed by silver staining. (D) Western blotting for YB-1, hnRNP L, hnRNP LL, hnRNP A1, and PCBP1 in the material pulled down by biotinylated scrambled or YB-1 RNA decoy oligonucleotides from U87 cell extracts. (E–G) Western blot analysis of YB-1, CCT4, mLST8, and mTOR markers in U251 (E), U87 (F), and GSCWL1 (G) cells transfected with scrambled or YB-1 decoy oligonucleotides. (H) Effects of scrambled or YB-1 decoy oligonucleotides on cell proliferation were tested in GSCWL1 cells. Data are presented as mean ± SEM (n = 3). ***P < 0.001 by unpaired, 2-tailed Student’s t test. (I) In vitro extreme limiting dilution assays were performed in GSCWL1 cells transfected with scrambled or YB-1 decoy oligonucleotides. (J) H&E-stained sections of tumor-bearing mouse brains. Tumors were formed by intracranial injection of GSCWL cells transfected with scrambled or YB-1 decoy oligonucleotides (scrambled, n = 8; YBX1-2, n = 7). Scale bar: 2 mm. (K) Bioluminescence images of tumor-bearing mouse brains described in J. Colored scale bar represents photons/s/cm2/steradian. (L) Total flux (photons/s) was determined by bioluminescence imaging (BLI) for the times indicated after intracranial injection of GSCWL1 cells described in J. Data are presented as mean ± SEM. ***P < 0.001 by 2-way ANOVA. (M) Kaplan-Meier survival curves of nude mice described in J. P value was determined by Mantel-Cox log-rank test. (N) Schematic illustration of the working model.