Figure 5.

Prolidase elicits migration in HaCaT cells through EGFR. (A) The level of expression of EGFR and Akt proteins analyzed by Western immunoblotting in lysates of scratched HaCaT cells incubated with PEPD (10 nM) and an inhibitor of EGFR (Gefitinib, 2 µM) for 30 min and 24 h. Representative blot images are shown (densitometric analysis of presented protein is included in Supplementary Materials (Supplementary Figure S4)). (B) Prolidase- and Gefitinib-treated keratinocytes were scratched and monitored using an inverted microscope at 0 and 24 h of incubation. Supplementary Materials contain microscopic images in triplicates (Supplementary Figure S5). (C) The wound closure rate of scratched HaCaT cells was evaluated by ImageJ software. The results represent mean ± SEM of three replicates and are significant aaaa, or cccc < 0.0001 and are marked as a vs. control (0 nM of PEPD) of scratched cells, b vs. control (0 nM of PEPD) of scratched cells after treated with Gefitinib, and c marked scratched cells vs. scratched cells after treated with Gefitinib in the same concentration of PEPD, respectively.