Fig. S10

Paclitaxel impairs skin wound repair. (A and B) Rapid repair of a puncture wound (dashed circles) in a vehicle-treated transgenic Tg(tp63:CAAX-GFP) zebrafish larva (A). The puncture wound is retained in a larva incubated in 22 µM paclitaxel (B). Animals were preincubated in 22 µM vehicle or paclitaxel for 3 h, injured, and retained in the treatment solution for 12 h. (Scale bar, 20 µm.) (C) Quantification of wound diameter over 12 h shows rescue of wound repair with MMP-13 inhibitors (n = 2, 4-5 larvae per group). (D, D′, and D′′) Scratch wounding of HEK001 human keratinocytes shows ROS/H₂O₂ formation at the scratch wound margin of vehicle (yellow arrows, D) but not paclitaxel-treated cells (D′ and D′′). (Scale bar, 50 µm.) (E and E′) ROS/H₂O₂ formation is present in vehicle (E) and paclitaxel-treated HEK001 cells (E′) after 12 h. (Scale bar, 50 µm.) (F and F′) Healed scratch in vehicle (F) but not paclitaxel-treated wells (F′) after 24 h. (G) ROS/H₂O₂ fluorescence intensity measured from the scratch margin 1 h postscratch (hps). (Scale bar, 100 µm.) (H) HEK001 scratch wound gap size over time (n = 5; ***P < 0.001). (I) HEK001 gap closure distance at 12 hps following Pctx + CL-82198 or CL-82198 alone administration. (J) HEK001 gap closure distance at 18 hps following Pctx + DB04760 or DB04760 alone administration. (K) Still images (Eosin) of HEK001 cells from J. Insets on the right of each image show higher magnifications of highlighted boxes. Control cells at the scratch margin show decreased cell-cell adhesion and formation of lamellipodia at the leading (migratory) edge, which is absent in paclitaxel-treated cells and rescued with DB04760. DB04760 alone increases the number of cells with lamellipodia formation. (Scale bar, 20 µm.) *P < 0.05, **P < 0.01, ***P < 0.001. a.u., arbitrary units; CL, CL-82198; DB, DB04760; hpp, hours postpuncture; hps, hours postscratch; mpp, minutes postpuncture; Pactx, paclitaxel; SC, scratch.