Fig. 8.

Hsa_circ_0058124 affects the NOTCH3 pathway via miR-218-5p/NUMB. a The interaction network between the NOTCH3 pathway and hsa_circ_0058124 using GeneMANIA web tool [31]. b Venn diagram showing the mutual putative target genes of hsa_circ_0058124 and NUMB predicted by CircInteractome and starBase. c Dual-luciferase assays showing the luciferase activity of the pmiR-RB-Report-hsa_circ_0058124 vector in 293 T cells cotransfected with indicated miRNA mimics. d Dual-luciferase assays confirmed the interaction of NUMB with miR-218-5p. e NUMB protein level after transfection with miR-NC or miR-218-5p. f, g mutant (mut) and wild-type (wt) luciferase reporter constructs of hsa_circ_0058124 (f) or NUMB (g) were transfected into TPC-1 cells together with miR-218-5p mimics or controls (miR-NC). Dual-luciferase assays were conducted to detect relative luciferase activity. h Expression level of miR-218-5p in TPC-1 and K1 cells infected with si-hsa_circ_0058124 or negative control (si-NC) were measured by qRT-PCR. i-j NUMB mRNA (i) and protein level (j) in TPC-1 and K1 cells transfected with si-hsa_circ_0058124 or si-NC. k Western blot assays were performed to measure the NOTCH3 and HES1 protein level in TPC-1 cells with indicated treatments. l-m CCK-8 assays (l) and transwell assays (m) in TPC-1 cells with indicated treatments