Fig 2: MiR-190a inhibits prostate tumor growth in vivo.(a–c) C4-2 tumors with stable overexpressed miR-190a injected into nude mice. Tumor sizes were measured every 5 days. The data is shown as mean ± SEM for N > 6 separate tumors for each group. (a) Images of tumors dissected from the mice. (b) The tumor sizes (mm3) versus days of post injection. (c) Tumors weights after resection at the end of experiment. (d–g) mRNA levels of miR-190a, AR, YB-1, CDKN1 (p21) determined by qRT-PCR from tumors. (h) IHC staining showing protein expression of AR, YB-1, Ki67, in C4-2 tumor tissues derived from mice. Data for quantified IHC was shown as mean ± SEM for N = 4 tumors in each group.

Fig 3: Upon knockdown of YBX1, PCBP1, and PCBP2 EVT-like cells fail to differentiate toward an invasive phenotype. (A) qPCR results demonstrating successful mRNA downregulation of PCBP1, PCBP2, and YBX1 in HTR-8/SVneo cells after transfection with respective siRNAs. An siRNA not targeting any known mammalian gene was used as a control. Normalization was done using the geometric mean of reference genes PSMD4 and YWHAZ. Experiments were repeated 3 times using 3 replicates per treatment. (B) Successful siRNA-induced downregulation of target genes on protein level as shown by Western blot. Equal protein loading control was performed by total protein Revert stain (Supplementary Figure 1). (C) Transwell invasion assays show that PCBP2 and YBX1 downregulation leads to a significant decrease in invasive behavior of EVT-like cells. Experiments were repeated 3 times using 6 replicates per treatment. (D) Quantification of proliferation using alamarBlueTM solution. The proliferation rate of EVT-like cells with downregulated expression of PCBP2 was increased. Experiments were repeated five times using nine replicates per treatment. Data are presented as mean ± SD and tested with two-way ANOVA followed by Dunnett’s multiple comparisons test or with a Student’s t-test in case of two data sets. *Indicates p < 0.05; *** indicates p < 0.001; **** indicates p < 0.0001.

Fig 4: Irradiation induces YB-1 expression in brain tumor initiating cells. (A) R28, R11 and R49 cells were irradiated with 2 Gy. Cell lysates were prepared at the indicated time periods (2-8 h) following irradiation. YB-1 expression was determined by western blot analysis. GAPDH was used as a control. (B) Upregulation of total YB-1 following irradiation at the indicated time points. YB-1, Y-box binding protein-1.

Fig 5: YB1 regulated ITGB8 to affect the RCC cell adhesion. Increased adhesion capability of 786-0-YB1 cells was rescued by adding RGD. The RGD peptide (Dalian Meilun Biotechnology Co. LTD) was dissolved in ddH2O at 10 mg/ml. 786-0-YB1 cells were incubated with 0.1 mg/ml RGD for 24 h before harvested. (* indicate p < 0.05, *** indicate p < 0.001)