Fig. 5

NovelmiRNA-25 targets AMPD2. a Predicted target sites in the AMPD2 3′UTR based on the NovelmiRNA-25 seed region. Mutations in the AMPD2 3′UTR generated missense sequences unable to pair with the NovelmiRNA-25 seed region. The structure of NovelmiRNA-25 bound to AMPD2 is depicted. b NovelmiRNA-25 and the AMPD2 3′UTR seeding region, highlighted in green, are highly conserved in mammals. c Activity of the luciferase gene linked to the 3′UTR of AMPD2. pmirGLO firefly luciferase reporter plasmids with wild-type or mutated 3′UTR sequences of AMPD2 were transiently transfected into cells with NovelmiRNA-25 precursor or negative control and a Renilla luciferase reporter for normalization. Luciferase activities were measured after 48 h. The mean of the results from the cells transfected with pmirGLO control vector was set as 100%. Data are the mean and standard deviation (SD) of separate transfections (n = 3). d Downregulation of endogenous AMPD2 protein expression by NovelmiRNA-25. Western blotting of AMPD2 protein after transfection with negative control (NC) or NovelmiRNA-25 mimic in HEK293T cells. Expression levels were normalized to that of GAPDH; **P < 0.05; AMDP2, adenosine monophosphate deaminase 2; e AMPD2 protein expression in PBMCs from SLE patients and negative control. Western blotting of AMPD2 protein after transfection with PBMCs. Expression levels were normalized to those of GAPDH. The arrow indicates the expression of AMPD2 protein