Long noncoding RNA MARL regulates antiviral responses through suppression miR-122-dependent MAVS downregulation in lower vertebrates
Fig 9
The ceRNA network of regulating MAVS is widely found in other species.
(A) Sequence alignment of pre–miR-122 from various vertebrate species. Mature miR-122 sequences are shown in boxes. (B) Putative miR-122-binding site of MAVS 3’UTR among different vertebrate species. (C) miR-122 target D. rerio MAVS 3’UTR and regulates its expression. ZFL cells were transfected with NC or miR-122, along with the wild-type of D. rerio MAVS 3’UTR (dreMAVS-3’UTR-wt) or the mutant-type (dreMAVS-3’UTR-mut). The luciferase activity was measured and normalized to renilla luciferase activity (upper panel). ZFL cells were transfected with NC or miR-122 for 48 h. MAVS expression were analyzed by western blotting (lower panel). (D) miR-122 target L. crocea MAVS 3’UTR and regulates its expression. LLC cells were transfected with NC or miR-122, along with the wild-type of L.crocea MAVS-3’UTR (lcrMAVS-3’UTR-wt) or the mutant-type (lcrMAVS-3’UTR-mut). The luciferase activity was measured and normalized to renilla luciferase activity (upper panel). LLC cells were transfected with NC or miR-122 for 48 h. MAVS expression were analyzed by western blotting (lower panel). (E) miR-122 target H. Sapiens MAVS 3’UTR and regulates its expression. HEK293 cells were transfected with NC or miR-122, along with the wild-type of H. sapiens MAVS-3’UTR (hsaMAVS-3’UTR-wt) or the mutant-type (hsaMAVS-3’UTR-mut). The luciferase activity was measured and normalized to renilla luciferase activity (upper panel). HEK293 cells were transfected with NC or miR-122 for 48 h. MAVS expression were analyzed by western blotting (lower panel). (F) The finding that miR-122 regulating the luciferase activity of MARL is also examined in N. diacanthus and L. crocea. NLC cells were transfected with NC or miR-122, together with the wild type or mutated type of N. diacanthus MARL, then luciferase activity was analyzed and normalized to renilla luciferase activity (left panel). LLC cells were transfected with NC or miR-122, together with the wild type or mutated type of as well as L. crocea MARL, then luciferase activity was analyzed and normalized to renilla luciferase activity (right panel). (G) MARL reduces miR-122 activity in N. diacanthus and L. crocea. NLC cells or LLC cells were respectively transfected with N. diacanthus or L. crocea MARL expression plasmid, together with miR-122 sensor, miR-122 and pcDNA3.1 vector for 48 h. The luciferase activity was analyzed and normalized to renilla luciferase activity. All data represented the mean ± SE from three independent triplicated experiments. *, p < 0.05.