S1 File - Epigenetic DNA Methylation Linked to Social Dominance

(Fig A). Aquaria for raising Never Been Dominant (NBD) males. 10 NBD males were raised from 2 weeks post-fertilization in community tanks with 4 large suppressor D males and 4 females. 2 spawning shelters were placed in the tank to limit the number of mating territories available. (Fig B). Experimental aquaria. Two size-matched males were injected and placed in middle experimental compartment with 3 smaller females. On either side were community compartments separated by clear barriers with 2 smaller males and 3 smaller females allowing behavioral interactions through transparent barriers. (Fig C). Data comparing control and L-Methionine injected animals. Averaged sum of daily territorial behaviors with non-territorial behaviors subtracted, comparing L-methionine (blue) and vehicle-injected animals (dashed). N = 8 for each group. (Fig D). Data comparing control and zebularine injected animals. Averaged sum of daily territorial with non-territorial behaviors subtracted, comparing zebularine (red) and vehicle-injected animals (dashed). N = 8 for each group. (Fig E). Brain staining for methylation. Coronal brain sections from A. burtoni male brains in the region of the pre-optic area stained with a monoclonal antibody specific for methylated cytosine residues for the four treatment groups (L-methionine and its controls; Zebularine and its controls. Top row (5mC); Middle row (Dapi counterstain); Bottom row (merged images).(Fig F). Comparison of fluorescence intensities for Fig E. Fluorescence intensity in 5-mC stained cells (N = 40) in POA brain sections from zebularine, methionine, and control injected animals. There was no statistically significant difference in flurorescence intensity between any of the treatment groups. (DOCX)