Supplementary Fig. S1 BCL2 5’flanking ctrl siRNA luc -753 +32 Tra2b siRNA ctrl siRNA mock luc Tra2b siRNA 2 4 Relative luciferase activity Supplementary Fig. S1　 Tra2β knockdown dose not affect the promoter activity of BCL2 After treatment with 10 nM Tra2β or ctrl siRNA for 24 h, HCT116 cells were transiently transfected with the luciferase plasmids containing the 5’-flank of the human BCL2 gene from -959 to + 107 bp for 24 h. Luciferase activities in these cells were measured using the Dual-Luciferase Reporter Assay System (Promega, Madison, WI). Values are means ± SD (n = 4). Tra2β knockdown did not affect the activity of the BCL2 promoter.

Supplementary Fig. S2 TRA2β1 mRNA is not a target of miR-204 b c 125 1.25 1.25 100 1.0 1.0 0.75 TRA2b1 expression levels (fold change) 0.75 TRA2b1 expression levels (fold change) 0.75 miR-204 expression levels (normalized to U6) 0.5 0.5 0.5 0.25 0.25 0.25 ctrl 10 nM 25 nM ctrl 25 nM 100 nM ctrl siRNA Tra2b siRNA pre-miR-204 anti-miR-204 Supplementary Fig. S2　 TRA2β1 mRNA is not a target of miR-204 (a, b) After HCT116 cells were treated with control (ctrl), precursor (pre)-miR-204, or anti-miR-204 at the indicated concentrations for 48 h, the amounts of Tra2β 1 and GAPDH mRNAs were determined by qPCR. The values are means ± SD (n = 5). (c) After transfection with ctrl or TRA2β siRNA for 24 h, the levels of mature were measured by qPCR. U6 snRNA was used as an endogenous quantity control. These results suggest that neither overexpression nor knockdown of miR-204 affects TRA2β1 mRNA levels, and that Tra2β knockdown dose not affect miR-204 levels.