Fig. 5

Human MCL-1 was targeted by shrimp miR-965 in a cross-species manner in MSLC. a Shrimp miR-965 was predicted to target human MCL-1 mRNA 3'UTR. b The interaction between shrimp miR-965 and human Ago2 protein. Shrimp miR-965 was incubated with recombinant human Ago2 protein. The mixture was separated by 1% agarose gel and stained with ethidium bromide to visualize the miRNA (top), followed by staining with Coomassie Blue (bottom). The wedges indicated the concentration gradient of the recombinant protein used. c The interaction between shrimp miR-965 and the MCL-1 mRNA in MSLC. Shrimp miR-965 or the control miRNA was transfected into MSLC. Twenty-four hours after transfection, the MCL-1 mRNA was detected using quantitative real-time PCR. d Western blot analysis of MCL-1 in shrimp miR-965-transfected MSLC. e The direct interaction between shrimp miR-965 and the MCL-1 gene. MSLC were co-transfected with shrimp miR-965 and a luciferase reporter fused with MCL-1 3′ UTR. At 24 h after transfection, the firefly and Renilla luciferase activities were analyzed. Control miRNA (shrimp miRNA-965-scrambled) and MCL-1 3′ UTR mutant were included in the co-transfections