Figure 3. Validation of hsa-miR-660-3p as the target of hsa_circ_0004018. (A, B) HSCs lysis was pulled down with hsa_circ_0004018 specific probe, and then hsa_circ_0004018 (A) and hsa-miR-660-3p (B) were respectively detected by real-time PCR. (C, D) The HSCs were transfected with biotin labeled hsa-miR-660-3p and the negative control (NC) following by strepavidin enrichment, and then hsa_circ_0004018 was detected by real-time PCR. (E) The sequences between hsa_circ_0004018 and hsa-miR-660-3p were compared, and the complementary bases of red color indicate the seed sequence of hsa-miR-660-3p. The mutant hsa_circ_0004018 was designed without the seed sequence. (F) The HSCs stably expressing the luciferase construct containing the wild-type (WT) or mutant (MT) hsa_circ_0004018 were respectively transfected with hsa-miR-660-3p mimic, inhibitor and the negative control, and then the luciferase activity was examined.