Fig. 3

Cell morphology, mRNA expression, WB results. (A) After endothelial cells transfected with overexpression of GDF15 plasmid, the mRNA expression of GDF15 was significantly up-regulated by RT-qCPR. (B) Western blot assay was used to detect GDF15 protein expression changes in GDF15 knockout cells and overexpressed cells, as well as endothelial and interstital marker changes after cytokine treatment, in which CON was the control group, CON + C was the control group + cytokine, Si was GDF15 knockout, OE was overexpressed GDF15. (C) After transfected with GDF15 plasmid and continued culture for 48 H, messtitial cell characteristics appeared in the morphology of endothelial cells, with a scale of 500 μm (40×). Cell scratch experiments (D, E) To reduce the influence of cell activity changes and proliferation on the scratch experiment results, we starved the cells for 24 H in advance, and used 1% serum during the experiment. The crawling ability of endothelial cells was enhanced after cytokine stimulation or transfection of the overexpressed plasmid, while the crawling ability of endothelial cells induced by cytokine stimulation was decreased by instantaneous knockout of GDF15, scale 500 μm (40×). * in Figure D represents comparison with CON group, significant post hoc effects were revealed by the Bonferroni post hoc test. ***p < 0.001, ****P < 0.0001