Fig. 6

Nuclear localization of YBX1 enhanced NLGN3 expression in GBM. A. TCGA database showed that the expression of YBX1 was positively correlated with that of NLGN3 at the mRNA level. B and C. U87MG and U251MG cells were infected with shYBX1#1 and shYBX1#2. Cells were collected for Western blot analysis (B) and RT-qPCR (C). Data presented as the mean ± SD of three independent experiments, ***P < 0.001, one-way ANOVA. D and E. U87MG and U251MG cells were infected with YBX1 plasmids. Cells were collected for Western blot analysis (D) and RT-qPCR (E). Data presented as the mean ± SD of three independent experiments, ***P < 0.001, one-way ANOVA. F and G. U87MG and U251MG cells were infected with shControl or shNLGN3 for 48 h. Then, cells were transfected with pcDNA3.1 or His-YBX1 as indicated. After 24 h, cells were harvested for Western blotting analysis (F) and RT-qPCR analysis (G). Data presented as the mean ± SD of three independent experiments, ***P < 0.001, one-way ANOVA. H. ChIP-Seq peaks were mapped on the UCSC genome browser. Data presented as the mean ± SD of three independent experiments, ***P < 0.001, ns not significant, one-way ANOVA. I. The DNA motif of HSF4 was prepared using JASPAR online tools. J. Relative quantification of ChIP-qPCR. All data are shown as the mean ± SD from three replicates. ***P < 0.001, unpaired Student’s t-test. K. DNA electrophoresis of the products from the ChIP assay