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. 2021 Jun 25;12(7):649. doi: 10.1038/s41419-021-03920-4

Fig. 5. TNF-α-induced EndMT is reversible.

Fig. 5

A, B TNF-α removal reverses the spindle-shaped phenotype induces by TNF-α (A) and TNF-α removal reverses the CD31 marker downregulation induced by TNF-α, restoring CD31 levels present in untreated controls (B). Reciprocal reversion is observed for the expression of the mesenchymal marker α-SMA. HMVECs were treated or not (−) for 72 h with 100 ng/ml of TNF-α then, cells were washed (+/−TNF-α) or not (+TNF-α) and cultured for various times. Scale bars, 50 µm (inset) or 400 µm. Protein expressions were analysed by western blot. C TNF-α induces a reduction in the formation of tubules in a matrigel-based tube-formation assay (+TNF-α), and a tubule restoration is observed from 168 h after removal of TNF-α (+/−TNF-α). Scale bars, 1000 µm. β-tubulin was used as loading Ctrl for western blot analysis. Tube-formation scores were measured for three fields. Similar results were obtained in three different experiments. Significant differences are indicated by solid lines (**P < 0.05, ***P < 0.005 by t test). HMVEC human microvascular endothelial cell, TNF-α tumour necrosis factor-α, α-SMA α-smooth muscle actin, Ctrl control.