Fig. 4
From: A somatic mutation in PIK3CD unravels a novel candidate gene for lymphatic malformation
The effects of PIK3CD mutation on the proliferation and migration of HUVECs. a Schematic diagram showing the construction of HUVECs with stable overexpression of PIK3CD. Adenovirus vectors expressing mutant or wild-type PIK3CD were constructed. Then, the two overexpression vectors or empty vector was transfected into 293 T cells to produce the virus. The three viruses were infected into HUVECs, resulting in MT-, WT- and Ctrl-HUVECs, respectively, for subsequent experiments. b The infection efficiencies of the Ctrl, WT-PIK3CD or MT-PIK3CD vectors was verified by GFP fluorescence imaging. c, d Cell proliferation of Ctrl-, WT- and MT-HUVECs was evaluated at 0, 24, 48 and 72 h after infection by the CCK8 assay (c) and bright-field microscopy (d). e In the wound healing assay, images of MT-, WT- and Ctrl-HUVECs were acquired at 0, 4, 8 and 12 h after scratching the cell monolayer. f The images acquired above were quantitatively analysed by measuring the area of the scratched region lacking cells, and the wound healing rate was calculated as follows: migration area (%) = (A0 – An)/A0 × 100, where A0 represents the area of initial wound area, An represents the remaining area of wound at the metering time point. Data is representative of three independent experiments and three-five visual fields were randomly selected from the results for quantitative analysis. *p < 0.05, **p < 0.01 and ***p < 0.001 compared with the Ctrl group (Student’s t test)