Fig. 4

Upregulation of LAP3 promoted cell cycle process of BMECs. a Detection of cell proliferation using CCK-8 method. b Effect of LAP3 on cell migration by cell scratch test. c LAP3 affects the cell cycle distribution of malignant transformed BMECs. The malignant transformed BMECs were treated with the LAP3 inhibitor bestatin, which was stained with propidium iodide (PI) and detected for DNA content and cell cycle distribution by flow cytometry. d Western blotting analyzed the effect of knock-down of LAP3 on cyclin A1, cyclin D1, and p27 expressions in malignant BMECs. Full-length blots are presented in Supplementary Fig. S9. e Overexpression of LAP3 promoted the cell cycle of normal BMECs. LAP3 was overexpressed using a plasmid in normal BMECs. Cells were stained with propidium iodide (PI) and detected for DNA content and cell cycle distribution by flow cytometry. f Western blotting analyzed the effect of overexpression of LAP3 on cyclin A1, cyclin D1, and p27 expressions in normal BMECs. VC, pLenti-CMV-GFP-Puro. pLAP3, pLenti-CMV-LAP3-GFP-Puro. Full-length blots are presented in Supplementary Fig. S10. The relative level of the target protein was estimated using densitometry and the ratio was calculated relative to the GAPDH control. Bars represent mean values with error bars to represent SD from three independent replicates. Differences between mean values were assessed by two-tailed Student’s t-test. *p < 0.05; **p < 0.01; ***p < 0.001, compared to control