Fig. 3

ROCK1 is a direct target of miR-335-5p. a Schematic diagram showing the subcloning of the predicted miR-335-5p binding site (position 472–478) of the ROCK1 3′-UTR into a psiCHECK-2 luciferase construct. Predicted duplex formation between miR-335-5p and the wild type or mutant miR-335-5p binding site is indicated. b-c Luciferase activity of the constructs containing the wild type or mutant ROCK1 3′-UTR reporter gene in A549 and SPC-A1 cells co-transfected with miR-NC or miR-335-5p. Relative Renilla luciferase activity was determined and normalized against the firefly luciferase activity. d-e qRT-PCR and western blot analysis of the expression of ROCK1 in cells transfected with miR-335-5p mimics. **P < 0.01; ***P < 0.001