Fig. 2.

CTDSPL is a direct target of miR-181 family members. a Alignment of the seed sites in the human 3’-UTR of CTDSPL gene. The mutated 3’-UTR of CTDSPL is underlined. b and (c) MUM2b cells were transfected with 40 nM miR-NC, miR-181a, -181b, -181c, and -181d or as-miR-181a, -181b, -181c and -181d. Overexpression or knockdown of miR-181 family inhibited or enhanced CTDSPL expression, respectively. The gray level was analyzed by histogram. d and (f) Overexpression or knockdown of miR-181 expression inhibited or enhanced the Renilla luciferase activity, respectively. 293 T cells were cotransfected with 40 nM miR-NC, miR-181a, -181b, -181c, and -181d, or as-miR-181a, -181b, -181c, and -181d, and 100 ng of reporter plasmid containing the wild-type 3’-UTR of CTDSPL. After 24 h, Renilla luciferase values, normalized against firefly luciferase, were measured. e and (g) The Renilla luciferase activity was nearly unchanged after mimics and inhibitors of miR-181 family members were transfected with the mutated 3’-UTR of CTDSPL