Fig. 1

BIX01294 treatment decreases H3K9me2 levels in zebrafish neuromasts (NMs). (A) Both the BIX01294 (BIX)-treated group and DMSO control group were morphologically normal following 1 h neomycin treatment. (B1,D1) H3K9me2 immunofluorescence (red) is of relatively high intensity in the control zebrafish NMs, and the H3K9me2 level increased at 48 h after neomycin damage for 1 h. (C1,E1) BIX01294 treatment reduced the H3K9me2 level, and the intensity of H3K9me2 was significantly decreased with the addition of 20 µM BIX01294 for 48 h following 1 h neomycin damage. (B2-E3) Some degree of decreased hair cell (HC; green) regeneration after neomycin treatment for 1 h was also observed after exposure to BIX01294 for 48 h compared to the neomycin treatment controls, while a reduction in HCs was not obvious in the non-neomycin control groups. HCs were labeled with anti-GFP antibody (B2-C2) or green fluorescence (D2-E2), and nuclei were labeled with 4,6-diamidino-2-phenylindole (DAPI; blue). Scale bar = 10 µm. (F) The reduced H3K9me2 level by 20 µM BIX01294 was reconfirmed with western blotting analysis at 48 h after neomycin damage. β-actin was included as the control. Mean ± SEM for three experimental replicates. **p < 0.01.