Fig. 4. miR-4521 downexpression accompanied with FAM129A overexpression in RCC cell lines through binding with the latter’s 3′-UTR domain.

a miR-4521 and FAM129A expressions in RCC cell lines (786-O and ACHN) compared with that in HK-2. b Sequence alignment of the FAM129A 3′UTR with wild-type (WT) versus mutant (MUT) potential miR-4521 targeting sites. c Dual-luciferase reporter assays showed decreased reporter activity after transfection of the wild-type FAM129A 3′UTR reporter construct in 786-O cells overexpressing miR-4521. The FAM129A 3′UTR MUT and control constructs showed no effect on reporter activity. d Alteration of the miR-4521 expression levels of 786-O and ACHN cells 24 h after transfection with the miR-4521 mimics or NC mimics. mRNA (e) and protein (f) levels of FAM129A were examined by qRT-PCR and Western in 786-O and ACHN cells that were transfected with miR-4521/NC mimics. *P values < 0.05, **0.01, ***0.001, ****0.0001