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. Author manuscript; available in PMC: 2017 Nov 1.
Published in final edited form as: Exp Eye Res. 2016 Sep 4;152:10–33. doi: 10.1016/j.exer.2016.08.020

Figure 5.

Figure 5

Glucocorticoids delay in vitro wound healing of HCE cells. Scratch assay was performed on confluent monolayers of cells pre-treated overnight alone or in combination with vehicle, DEX (1000nM), or RU486 (10uM) in charcoal-stripped serum containing medium. Real-time analysis of cell migration was performed by imaging the every 30 minutes for 18 hours post-scratch. A) Representative images of wound healing kinetics in all the 4 conditions-vehicle, dexamethasone (1000nM), combination of dexamethasone (1000nM) and RU486 (10uM) and RU 486 (10uM) alone. Scratch width and time are on the X- and Y- axes, respectively. Yellow dots indicate the edge of the scratched monolayer. B) Representative images showing the time-projection of wound healing over a period of 18 hours in all four conditions. Time-projection is indicated with t0 in white and t18hrs in red. C) Quantification of the extent of wound healing measured from time-lapse images taken over a period of 30 hours. Average of four individual experiments is represented here (*p<0.001). D) Average area of wound closure in 18 hours represented in percentage from four individual experiments (*p<0.05). E) Representative images showing the net change in the area of the lamellipodia in vehicle and dexamethasone (1000nM) treated cells at 30 minutes after scratching the monolayer. Red represents lamellipodia that are moving forward to close the wound, green represents retraction of the lamellipodia and yellow represents no change in net movement of the lamellipodia over a period of approximately 6 minutes. Arrows are pointing to the filopodia. F) Average of the change in the area of the lamellipodia per minute in 10 minutes after creating a scratch wound in the monolayer in HCE cells treated overnight with either vehicle or 1000nM dexamethasone. Results from three experiments were averaged and are shown here. G) Average number of filopodia formed at the wounded monolayer between 10-65 minutes of wound healing. An average of three individual experiments is represented here.