Fig. 7: Melatonin regulates EMT through the GDF15-SMAD3 pathway to promote trophoblast cell transplantation and migration. | Communications Biology

Fig. 7: Melatonin regulates EMT through the GDF15-SMAD3 pathway to promote trophoblast cell transplantation and migration.

From: Melatonin affects trophoblast epithelial-to-mesenchymal transition and oxidative damage resistance by modulating GDF15 expression to promote embryo implantation

Fig. 7: Melatonin regulates EMT through the GDF15-SMAD3 pathway to promote trophoblast cell transplantation and migration.

After HTR-8/SVneo cells treated with different concentrations of melatonin for 24 h, the protein expression of GDF15, SMAD1, SMAD5, SMAD2/3, p-SMAD3, p-SMAD1/5, E-CAD, and N-CAD was detected (A). The protein expression of GDF15, SMAD2/3, p-SMAD3, E-CAD and N-CAD was detected after GDF15 knockdown treatment (B). After knocking down the expression of GDF15, melatonin (10 μM) was added again for 24 h to detect the protein expression of GDF15, SMAD2/3, p-SMAD3, E-CAD, and N-CAD (C). SIS3 (5 μM) was used for 6 h to inhibit SMAD3 protein phosphorylation and then add melatonin (10 μM) again for another 24 h to detect the protein expression of SMAD2/3, p-smad3, E-CAD and N-CAD (D). B-ACTIN/GAPDH is the internal reference protein. HTR-8/SVneo was cultured in a low-attachment 96-well plate for 24 h to form cell spheroids and co-cultured with ishikawa (E), and analyze the number of trophoblast cell spheres planted in each group at 0.5 h and 2 h (F, G). Transwell migration assay was used to detect the migration ability of cells in each group (H), and statistically analyzed the number of migrating cells in each group (I, J). Scale bar is 50 μm. Ns indicates no statistical significance, * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001, **** indicates p < 0.0001.

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