Non targeting control oligos

Synthetic siRNA oligos targeting hsYY1 and non-targeting control oligos were obtained from Guangzhou Ribo Bio Co. Ltd. The transfection of 293 T cells was performed with the indicated siRNA using Lipofectamine^® RNAiMAX according to the manual (cat.: 11668-019, Thermo Fisher Scientific). Cells were harvested at 48 h post-transfection. The hsYY1 knockdown efficiency was analyzed by western blotting (using anti-YY1 (H414; sc-1703) or anti-YY1 (C20; sc-281) antibodies, Santa Cruz). Signals were normalized to GAPDH.
For 293T^shYY1 cell construction, three pairs of hsYY1 shRNA oligomers were synthesized and annealed according to the manual’s instructions (Thermo Fisher Scientific). The lentiviral pLKO.1-shYY1 plasmids were first constructed. Viral particles were then prepared by transfecting psPAX2 and pMD2.G with pLKO.1-shYY1 plasmids into 293T cells. A more detailed description of the lentiviral package can be obtained from the instructions for Addgene Plasmid 10878. Finally, the constructed 293T^shYY1 cells were screened out via puromycin resistance and maintained as usual with puromycin (2 µg mL⁻¹) addition. The hsYY1 knockdown efficiency was confirmed by western blotting. Three pairs of hsYY1 shRNA oligomers are listed in Supplementary Table 1.

The human miR-139 mimic oligos, miR-139 inhibitor, and non-targeting control oligos were obtained from RiboBio (Guangzhou, China). The mimics, inhibitors and control oligos were transiently transfected into Hep3B and SMMC-7721 cells at a final concentration of 50 nM by using lipofectamine 2000 (Thermo Fisher Scientific, USA) according to the manufacturer’s instructions.