Fig. 6

Hsa_circ_0002577 regulates EC cell proliferation and migration via the modulation of IGF1R and PI3K/Akt signaling pathway. Ishikawa cells were co-transfected with sh-circRNA (or sh-NC) and lentiviral vectors expressing IGF1R (or control vectors). a Cell proliferation at indicated time points (0, 24, 48, 72, and 96 h) post-transfection was evaluated using CCK-8 assay. b The number of colonies was measured 10 days after transfection. c The apoptosis of transfected Ishikawa cells was detected using TUNEL and DAPI staining. The percentage of TUNEL-positive cells was calculated. The migration capability of cells after transfection was assessed by d wound healing assay and e Transwell assay. f HUVECs were cultured with ECGM containing the supernatants collected from Ishikawa cells co-transfected with sh-circRNA (or sh-NC) and lentiviral vectors expressing IGF1R (or control vectors) for 4 days. Then HUVECs were transferred to Matrigel-coated wells and incubated at 37 °C for 24 h. The number of branches in each group of cells was quantified. g The protein expressions of IGF1R, p-Akt, Akt, PCNA, p27, Bcl-2, cleaved caspase-3, E-cadherin, and N-cadherin were measured by Western blot and normalized to GAPDH