Fig. 5

Identification of MAP1LC3B as a novel direct target for miR-342-3p. a By qRT-PCR, in SU-DHL-16 cells, the expression of MAP1LC3B was downregulated upon overexpression of miR-342-3p compared with negative scramble control. Data were normalized to negative scramble control as 1. b By qRT-PCR, the expression of MAP1LC3B was significantly higher in NHL cell lines methylated for EVL/MIR342. c Sequences of putative miR-342-3p binding sites in cloned fragment of MAP1LC3B 3′-UTR, showing one bioinformatically predicted putative seed region binding site (site 1) and two 5-mer binding sites (sites 2 and 3). The deletion mutant was generated according to the bottom line of each binding site. d Luciferase plasmids containing wild-type or mutant MAP1LC3B 3′-UTR were co-transfected with miR-342-3p mimics or scramble control into HeLa cells. Luciferase reporter assay was performed at 48 hours post-transfection. Co-transfection of both miR-342-3p mimics and wild-type MAP1LC3B 3′-UTR significantly reduced luciferase assay signals. Deletion of the putative SRBS did not restore the luciferase signal whereas deletion of all three binding sites could restore the luciferase activity as compared with wild-type MAP1LC3B 3′-UTR. Data were normalized to negative scramble control as 100%. Columns represented mean +/− 1SD from three experiments in triplicate