Fig. 7

ADAMTS9-AS1 guides HuR by forming an RNA–RNA complex with ADAMTS9 3’UTR. A. The binding sites of ADAMTS9-AS1 and ADAMTS9 3’UTR were predicted by website (IntaRNA). B. Schematic of the predicted positions of ADAMTS9 3’UTR binding sites in ADAMTS9-AS1 (left). RNA–RNA interaction assay was used to detect the binding of biotin-labeled full-length or truncated ADAMTS9-AS1 and ADAMTS9 3’UTR in A498 cells (right). The same amount of biotin-full-length or truncated ADAMTS9-AS1 transcripts confirmed by dot-blot assay. C. Schematic of the predicted positions of ADAMTS9-AS1 binding sites in ADAMTS9 3’UTR (left). RNA–RNA interaction assay was used to detect the binding of biotin-labeled full- length or truncated ADAMTS9 3’UTR and ADAMTS9-AS1 in A498 cells(right). The same amount of biotin-full-length or truncated ADAMTS9 3’UTR transcripts confirmed by dot-blot assay. D. RNA–RNA interaction assay was used to determine the biotin-labeled ADAMTS9-AS1#3 (551–882 nt) directly binding with full-length or truncated ADAMTS9 3’UTR in vitro. The same amount of biotin-full-ADAMTS9-AS1#3 transcripts confirmed by dot-blot assay. E. Full-length and truncated ADAMTS9 3’UTR luciferase activities were measured by luciferase reporter assay in ADAMTS9-AS1 overexpressed A498 cells. The data derived from three independent experiments are presented as the mean ± SEM in the bar graphs (B–E). Controls were normalized to 1 (E). *P < 0.05, **P < 0.01, N.S. not significant