Fig. 4

Inhibition of macrophages has no significant effect on the differentiation of regenerating neuromasts. (a) At 3 days post-fertilization, we performed tail amputation on Et(HG7L) larvae. At 2 days post-amputation (dpa), larvae were either left untreated (Unt) or injected with 2 nL of the control encapsome (En) or clodrosomes (Clo) into the posterior cardinal vein. The percentage of surviving larvae at each phase for each treatment was recorded at the designated times, as outlined in Figure 1. The chi-square test did not identify significant differences in the ratios of phases between groups for each time point. For each treatment, n = 56–59 samples were used, and the inclusion or exclusion of deceased larvae did not alter the results of statistical analyses. At the designated time points, larvae were labeled with Rhodamine 123 (R123) to visualize live hair cells. An illustration of an untreated larva at Phase III with R123-labeled hair cells is provided in (b). Subsequently, the hair cell-labeled larvae underwent immunohistochemistry against Sox2 to highlight mantle cells (MCs) and supporting cells (SCs). An untreated larva at Phase III with Sox2-expressing cells in the regenerating neuromast is shown in (c), with a magnified view on the right. (d, e) Grouped column scatter plots with error bars display the numbers of R123-positive (R123+) cells (d) and Sox2+ cells (e) per neuromast (the regenerating neuromast). The number of hair cells differed significantly only between different time points (ANOVA, p < .01), and no significant effects were observed for the number of Sox2+ cells. Error bars represent SEM.