Fig. 1

SOX9 is a direct target of RUNX2 in OS. A, Venn diagram showing the strategy of identifying transcription factors regulated by RUNX2. RUNX2-regulated transcripts were derived from public datasets (GSE76937 and GSE77352). The GO term for transcription factors is 0003700. B, Heatmap showing the fold change (FC) of shLuc versus shRUNX2_3 or shRUNX2_4 for the eight transcription factors regulated by RUNX2. C, RNAseq showing FPKM (fragment per kilobase per million reads) of SOX9. The RNAseq data is from a public dataset, GSE77352. D, Immunoblotting showing the effect of RUNX2 knockdown of SOX9 in SAOS2 cells. E, ChIPseq showing the binding of RUNX2 on the SOX9 locus in SAOS2 cells. The ChIPseq data is from a public dataset, GSE76937. The putative RUNX2 response element (RE) was shown in red, and the DNA sequence comprising this RE was cloned to a reporter, pGL4.23 (See Methods for details). F, Reporter assays in SAOS2 (left) and 293 T (right) cells showing that the putative RUNX2 response element (RUNX2_RE) is involved in the regulation of SOX9 by RUNX2. No_RE: empty pGL4.23 vector (No response element); EV: empty vector (without RUNX2 overexpression). n = 4, p values are from the t-test