The goldenrod gall fly (Eurosta solidaginis)is a well-studied model of insect freeze tolerance. In situations of prolonged winter subzero temperatures, larvae ofE. solidaginisaccept ice penetration throughout extracellular spaces while protecting the intracellular environment by producing extreme amounts of glycerol and sorbitol as cryoprotectants. Hypometabolism (diapause) is implemented, and energy use is reprioritized to essential pathways. Gene transcription is one energy-expensive process likely suppressed over the winter, in part, due to epigenetic controls. The present study profiled the prevalence of 24 histone H3/H4 modifications ofE. solidaginislarvae after 3-week acclimations to decreasing environmental temperatures (5 °C, −5 °C and −15 °C). Using immunoblotting, the data show freeze-mediated reductions (p< 0.05) in seven permissive histone modifications (H3K27me1, H4K20me1, H3K9ac, H3K14ac, H3K27ac, H4K8ac, H3R26me2a). Along with the maintenance of various repressive marks, the data are indicative of a suppressed transcriptional state at subzero temperatures. Elevated nuclear levels of histone H4, but not histone H3, were also observed in response to both cold and freeze acclimation. Together, the present study provides evidence for epigenetic-mediated transcriptional suppression in support of the winter diapause state and freeze tolerance ofE. solidaginis.