Fig. 3

Forskolin treatment reduces iridoblast marker pnp4a expression. (A–C) Lateral brightfield images of larvae at 2 dpf processed for in situ hybridization using pnp4a probes, following treatment with 0.1% DMSO (A), 5 µM forskolin (B), or 25 µM H-89 (C) in embryo media. Activation of adenylyl cyclase (and PKA signaling) reduces the levels of pnp4a expression, whereas inhibition of PKA has the converse effect (increased pnp4a expression). Scale bar = 200 um andapplies to all images. (C’) Representative image with black arrows showing blue/purple pnp4+ cells included in panel D quantification. Brown melanophores were not included in counts. (D) Quantification of pnp4a+ cells confirms a significant reduction and increase in the number of pnp4a+ cells following forskolin and H-89 treatment, respectively (** p < 0.01 and *** p < 0.0001 via two-way ANOVA and Bonferonni multiple comparison analysis). (D’) The quantified regions are labeled.