Fig. 4
From: GDF15 affects venous thrombosis by promoting EndMT through smad2/p-smad2 pathway
Cell mRNA expression, WB and double luciflucidase assay. (A) After stimulating endothelial cells with TGF-β1 + TNF-α + IL-1β for 24 h, RT-qPCR was used to detect mRNA levels of related proteins. TF and PAI-1 expressions were up-regulated, while TFPI and PLAU expression levels were down. (B) After 24 h stimulation of endothelial cells by cytokine mixture, TF and PAI-1 expression decreased significantly after GDF15 knockout, *compared with CON group, # compared with CON + cytokine group. (C) Western-blot results, where CON is the control group and CON + C is the control + cytokine. After cytokine stimulation, Smad pathway was activated in endothelial cells. Knocking down GDF15 inhibited the phosphorylation of Smad2 and the expression of the transcription factor snail, while TF expression decreased significantly. (D) Dual luciferase reporter gene, SBE4 transcription level is up-regulated after EndMT, and knocking down GDF15 can significantly inhibit this effect. (E, F) Results of p-smad2 immunohistochemical staining of mouse inferior vena cava tissue, the scale was 200 μm (100×). Average optical density value of the positive signal of immunohistochemistry analyzed by Image J. Significant post hoc effects were revealed by the Bonferroni post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001