Fig. 3
LncRNA PAXIP1-AS1 regulates the expression of KIF14 by recruiting the transcription factor ETS1. a, Results of bioinformatics predictions. b, KIF14 expression in the glioma-related microarray dataset GSE104291. c, Western blot analysis of KIF14 expression normalized to GAPDH in glioma tissues, * p < 0.05 vs. the normal brain tissues. D, Western blot analysis of KIF14 expression normalized to GAPDH in TJ905 cells, * p < 0.05 vs. HAs. E, Pearson’s correlation analysis between lncRNA PAXIP1-AS1 and KIF14 expression in glioma tissues (n = 44). f, The dual luciferase reporter gene assay to examine the effect of lncRNA PAXIP1-AS1 on the KIF14 promoter activity, * p < 0.05 vs. the oe-NC group, # p < 0.05 vs. the sh-NC group. g, RIP assay verified that lncRNA PAXIP1-AS1 could bind to transcription factor ETS1, * p < 0.05 vs. IgG. h, The two sites of the KIF14 DNA promoter region that transcription factor ETS1 most likely to bind to. i, The truncated KIF14 recombinant luciferase reporter vector and ETS1 expression vector were constructed for transfection into TJ905 cells for dual luciferase reporter gene assay, * p < 0.05 vs. the oe-NC group. j, The mutant KIF14 recombinant luciferase reporter vector and ETS1 expression vector were co-transfected into TJ905 cells for dual luciferase reporter gene assay, * p < 0.05 vs. the oe-NC group. k, ChIP assay for the binding ability of ETS1 at binding site 2 of KIF14 promoter region, * p < 0.05 vs. IgG. l, ChIP assay for the enrichment of KIF14 by ETS1 after silencing lncRNA PAXIP1-AS1 in TJ905 cells, # p < 0.05 vs. the sh-NC group. m, Western blot analysis for the expression of KIF14 and ETS1 normalized to GAPDH in each group, * p < 0.05 vs. the oe-NC + sh-NC group, # p < 0.05 vs. the PAXIP1-AS1 + sh-NC group. The values are measurement data and expressed as mean ± standard deviation. Comparison of normally distributed data between two paired groups with homogeneity of variance was performed using a paired t test. The unpaired t test was used to compare two sets of data from independent groups with normal distribution and homogeneity of variance. Data comparisons between multiple groups were performed using one-way ANOVA with Tukey’s post hoc test. Experiments were repeated three times independently