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. 2019 Dec 23;8:e50047. doi: 10.7554/eLife.50047

Figure 7. BMP signaling regulates expression of and acts through mitfa to impact pigment cell fates.

(A) mitfa expression in sorted GFP-positive cells from Tg(crestin:eGFP) and Tg(mitfa:eGFP) embryos treated with vehicle or BMPi from 12 to 24 HPF, n = 4–5 replicates for each condition from two independent experiments (N = 2). (B) anti-Mitfa immunofluorescence, DAPI and merged images of Tg(crestin:eGFP) embryos treated with vehicle control or BMPi in GFP-positive cells (top) and GFP-negative cells (bottom), scale bar = 5 µm. (C) Quantification of anti-Mitfa fluorescence intensity of individual nuclei in GFP-positive cells (top) and GFP-negative cells (bottom); n = 65 and 74 for GFP-positive vehicle and BMPi groups, respectively; n = 35 and 30 for GFP-negative vehicle and BMPi groups, respectively, from three independent experiments (N = 3). (D) Incident light images of mitfa(lf) embryonic zebrafish treated with vehicle or BMPi from 12 to 24 HPF and imaged at 5 DPF, scale bar = 1 mm. (E) Quantification of dorsal iridophores in mitfa(lf) embryonic zebrafish treated with vehicle or BMPi from 12 to 24 HPF, n = 7 and 9 for vehicle and BMPi groups, respectively, from two independent experiments (N = 2). Error bars represent mean + /- SEM, P-value was calculated using one-way ANOVA with Tukey’s multiple comparisons test in panel A and Student’s t-test in panel C and E. *p<0.05, ***p<0.001, ****p<0.0001, n.s., not significant.

Figure 7—source data 1. BMP signaling regulates expression of and acts through mitfa to impact pigment cell fates.

Figure 7.

Figure 7—figure supplement 1. Diagram of synteny between MITF and mitfa loci and SMAD binding motifs at the mitfa locus.

Figure 7—figure supplement 1.

(A) Diagram of orthologous genes flanking the MITF and mitfa loci in human and zebrafish genomes, respectively. H. sapiens, Homo sapiens; D. rerio, Danio rerio. (B) Diagram of mitfa locus, including promoter, intronic and exonic regions with identified SMAD binding motifs (SBM) and GC-rich SMAD binding motifs (GC-SBM). Scale bar = 200 bp.