Figure 4. ANRIL-miR-181a-HMGB1 axis is critical for the progression pancreatic cancer. (A) The image reflected the binding sequence between miR-181a and HMGB1, and the corresponding mutant sequence between them. (B) Relative double luciferase activity experiment reflected the regulation of wild type HMGB1 and mutant HMGB1 activity by miR-181a. (C, D) Western blot and qRT-PCR analyzed the HMGB1 expression level which caused by miR-181a. (E) The image reflected the binding sequence of miR-181a and ANRIL, and mutant sequence between them. (F) Relative double luciferase activity experiment reflected the regulation of wild type ANRIL and mutant ANRIL activity by miR-181a. (G) The results of ANRIL mRNA and protein level indicated that it was regulated by miR-181a. *P < 0.05, **P < 0.01, ***P < 0.001.