ETS1 and SP1 are the direct targets of miR-324-5p.

<p><b>A.</b> Schematic representation of the mature miR-324-5p sequence, miR-324-5p target site in the 3'-UTR of <i>ETS1</i> and <i>SP1</i> mRNA and a 3'-UTR mutant of <i>ETS1</i> and <i>SP1</i> mRNA containing three altered nucleotides in the putative target site (shown as <i>ETS1</i> or <i>SP1</i>-3'UTR-mu). <b>B.</b> The expression levels of ETS1 and SP1 protein in HCC cells overexpressing miR-324-5p or transfected with miR-324-5p inhibitor, compared with control cells, by western blotting 48 hours after transfection; α-Tubulin served as the loading control. <i>C.</i> Luciferase assay of pGL3- <i>ETS1</i>-3'UTR or pGL3- <i>ETS1</i>-3'UTR-mut reporter, pGL3- <i>SP1</i>-3'UTR or pGL3- <i>SP1</i>-3'UTR-mut reporter cotransfected with different amounts (25, 50nM) of miR-324-5p mimic in indicated cells, or different amounts (50, 100nM) of miR-324-5p inhibitor, compared with negative control (NC). Each bar represents the mean ± SD of three independent experiments. * <i>P</i> <0.05.