Western painted turtle miR-29b and response to freezing. (a) Nucleotide sequence and predicted secondary structure of pre-miR-29b transcripts from H. sapiens, A. spinifera, and C. p. bellii at 25 C. Nucleotide substitution which leads to differential terminal stem-loop formation that is unique to C. p. bellii is circled. (b) Relative expression levels of miR-29b as assessed by quantitative RT-PCR in liver samples of hatchling western painted turtles under control (5°C acclimated), 24 h frozen (at -2.5°C), or 4 h thawed (at 5°C) conditions. Data are means ± s.e.m. (n = 5 different animals). Parallel analysis of 5S rRNA found no significant changes between control and experimental conditions for this reference RNA. * Significantly different from the corresponding control (P <0.05).