Fig. 6

Fig. 6

Qx protects against ototoxin-induced HC death and promotes supporting cell proliferation. (A?I) TUNEL assay (red) was performed in zebrafish incubated with vehicle or 300?µM of Qx alone (controls) or with the corresponding ototoxin with or without Qx 300?µM. Animals were counterstained with phalloidin (green). (A) Neomycin (Neo) 200?µM incubation for 30?min. (B) Gentamicin (GM) 50?µM incubation for 1 hour. (C) Cisplatin (CP) 400?µM incubation for 2 hours. (D) GM 50?µM incubation for 1 hour followed by recovery for 5 hours. (E) Incubation with Qx 300?µM for 8 hours and Neo 200?µM for 30?min. (F) Incubation with Qx 300?µM for 8 hours and GM 50?µM for 1 hour. (G) Qx 300?µM for 8 hours?+?CP 400?µM for 2 hours. (H) Qx 300?µM incubation for 8 hours?+?GM 50?µM for 1 hour followed by 5 hours recovery. Asterisks denote TUNEL-positive HCs. (I) The percentage of TUNEL-positive neuromasts was calculated for each treatment and represented as mean +/? SEM. (J?R) Proliferation assays were performed in 5dpf Tg(brn3c:GFP) in the presence/absence of Qx and the corresponding ototoxin, by the BrdU-labelling method (red). Animals were immunostained for GFP (green). (J) control. (K) Neo 200?µM 30?min. (L) GM 50?µM 1 hour. (M) CP: 400?µM 2 hours. (N) Qx 300?µM 8 hours. (O) Qx 300?µM 8 hours?+?Neo 200?µM 30?min. (P) Qx 300?µM 8 hours?+?GM 50?µM 1 hour. (Q) Qx 300?µM 8 hours?+?CP 400?µM 2 hours. Asterisks denote neuromast supporting cells positive for BrdU. (R) The percentage of BrdU-positive supporting cells per neuromast was calculated for each treatment and represented as mean +/? SEM. One-way ANOVA, *p?<?0.05, **p?<?0.01. Black asterisks compared versus corresponding control. Red asterisk compared versus the corresponding ototoxin-only treatment. Scale bar: (A?H) 10?µm, (J?O) 7??m. Data were taken from at least 15 animals and 3 experiments runs.