Fig. 3

MYLK-AS1 is a ceRNA of miR-424-5p in HCC. a MYLK-AS1 (red) detection in MHCC-97H cells by FISH. The nucleus was counterstained with DAPI (blue). b Relative MYLK-AS1 expression in six HCC cell lines by qRT-PCR. c Schematic representation of the predicted binding site for miR-424-5p in MYLK-AS1 by the online database Mircode Predicted algorithm. The mutated site in the 3′-UTR of MYLK-AS1 is shown as MYLK-AS1-Mut. The numbers indicate the positions of the nucleotides in the reference wild-type sequence of MYLK-AS1 (Ensembl version: ENSG00000239523). d-e Schematic representation of the predicted miR-424-5p target site within the 3′-UTR of E2F7. The predicted target site for miR-424-5p is located at the proximal portion of the E2F7 3′-UTR. Two nucleotides complementary to the seed sequence of miR-424-5p were mutated in the E2F7 mutant plasmid. The number indicates the position of the nucleotides in the reference wild-type sequence of E2F7 (NM_203394.3). f Relative miR-424-5p expression in Hep-G2 and MHCC-97H cells transfected with miR-424-5p mimic or inhibitor. g Relative miR-424-5p expression in Hep-G2 and MHCC-97H cells after transfection with shMYLK-AS1 or scramble sequence. h Relative MYLK-AS1 expression in Hep-G2 and MHCC-97H cells transfected with miR-424-5p mimic or inhibitor. i Correlation analysis between MYLK-AS1 and miR-424-5p expression in 156 HCC tissues. j Relative luciferase activity of the wild type (WT) and mutated (Mut) MYLK-AS1 reporter plasmid in human embryonic kidney (HEK) 293FT cells transfected with miR-424-5p mimic. Results are expressed as mean ± SD. n.s, not significant, *P < 0.05, **P < 0.01, and ***P < 0.001