Molecular Regulation of RUNX1-IT1 Expression

pcDNA3.1-control (vector) and pcDNA3.1-RUNX1-IT1 (RUNX1-IT1) were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China) as previously reported^{24 (link)}. Scrambled shRNA (shControl) and RUNX1-IT1 shRNAs (shRNA#1 and shRNA#2) were obtained from Invitrogen (Carlsbad, CA, USA). MiR-632 inhibitors and miR-632 mimics were synthesised from Genecopoeia (Guangzhou, China). The siRNAs against GSK-3β, HIF-1α and HDAC3 were designed and synthesised by Shanghai GenePharma Co., Ltd. (Shanghai, China). The sequences were as follows: si-GSK-3β/sense 5′-CGAUUACACGUCUAGUAUA-3′ and si-GSK-3β/antisense 5′- UAACAAUCUAUUUACACCC-3′; si-HDAC3/sense 5′-UCGCCUGGCAUUGACCCAUTT-3′ and si-HDAC3/antisense 5′-AUGGGUCAAUGCCAGGCGATT-3′; si-HIF-1α/sense 5′-UGUGAGUUCGCAUCUUGAUTT-3′ and si-HIF-1α/antisense 5′-TTACACUCAAGCGUAGAACUA-3′. The GSK-3β Human cDNA ORF Clone (GSK-3β) was purchased from OriGene (OriGene Technologies, Inc., USA). Cell transfection was conducted as previously described^{25 (link)}. Forty-eight hours post transfection, the cells were utilised for further experiments.