Fig. 4

DHA altered cell attachment, spreading, and focal adhesions. a The morphologies of GBC-SD and OCUG-1 cells that were treated with DHA were examined under a microscope. b GBC-SD and OCUG-1 cells were treated with DHA for 2 days. They were then re-plated on fibronectin-coated coverslips for 6 h before being washed, fixed, and stained with rhodamine-conjugated phalloidin. The attached cells were counted using light microscopy. c GBC-SD and OCUG-1 cells were processed and stained as described in (b). Images of the cells were taken using a confocal microscope, and cell sizes were measured using Volocity software. At least 50 cells were measured for each time group. d Representative images of focal adhesions in GBC-SD and OCUG-1 cells that were treated with DHA. Cells were re-plated on fibronectin-coated coverslips, fixed, and stained with anti-vinculin antibodies and rhodamine-conjugated phalloidin. The cells were visualized using a confocal microscope. e Total cellular protein levels were detected using western blot analysis with antibodies against vinculin and E-cadherin. β-actin was used as an internal control