Fig. 4

Activation of α7nAChR partially reverses platelet-induced EMT in endometriotic epithelial cells. A Relative fold change in gene expression levels of vimentin, SLUG and PAI-1 in 11Z cells treated with buffer, platelets activated with or without PNU-282987, or PNU-282987 alone for 48 h (n = 4). Values are normalized to GAPDH expression. B Left panel: Detection of protein levels of E-cadherin and α-SMA by immunoblotting of lysate of 11Z cells treated as in (A). Right panel: Relative fold changes of the protein levels of E-cadherin and α-SMA (n = 3). C Migratory capacity, as evaluated by the scratch assay, of 11Z cells treated with buffer, platelets with or without PNU-282987, or PNU-282987 alone for 12 and 24 h. The cells were photographed at 0 h, 12 and 24 h after scratch (n = 4). Fold change in migration distance was shown for different treatments as compared with controls (buffer). D Representative images of the invaded 11Z cells in the transwell assay under treatment indicated (Magnification: X200). Cells were added to the top of transwells coated with Matrigel and treated with buffer (Con), platelets with or without PNU-282987 (PLT, PLT + PNU), or PNU-282987 alone (PNU) for 48 h (n = 3). The total number of cells invaded to the bottom of transwell was then counted. Scale bar = 100 μm. E Representative immunofluorescence staining results of E-cadherin, α-SMA and F-actin expression in 11Z cells under different treatments as indicated. Symbols for statistical significance levels: NS: p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001