Fig. 4
From: Molecular features and vulnerabilities of recurrent chordomas
The HOX/PBX network is dysregulated by diminished miR-196a-5p levels in recurrent chordoma cell lines. a Scheme of the HOX clusters and miRNAs of the miR-196 family located within the cluster. Micro-RNA-196a-5p expression levels of matched (b) and unmatched (c) primary and recurrent chordoma cell lines. Relative fold change levels are given on the y-axis. (d) miR-196a-5p target sites (underlined nucleotides) in the 3’UTRs of HOXA7 and HOXB7 predicted by TargetScan 7.2. (e) Response to miR-196a-5p in U-CH1 cells. Firefly luciferase reporters containing either the complementary site of HOXA7 and HOXB8 or the perfect antisense sequence of miR-196a were co-transfected with a miR-196a mimic or a scrambled control, respectively. Firefly luciferase activity was normalized to Renilla luciferase activity. (f) Overexpression of miR-196a-5p in U-CH1 cells confirmed by qRT-PCR. (g) The effect of miR-196a-5p on HOXA7 levels in U-CH1 cells assessed by Western blot analysis. Experiments were performed in biological triplicates. T-tests were performed to determine statistical significance (*p < 0.05, **p < 0.01, ***p < 0.001). (h) GO-term analysis of the predicted target genes (n = 50) illustrated as cnetplot. (i) STRING analysis of the predicted miR-196a-5p target genes (n = 50) revealed an interaction network between HOX transcription factors and PBX co-factors