Differentiation-induced changes in Tfam mRNA stability (A) The nt sequence corresponding to the 3′-UTR of mouse Tfam is illustrated. Differentiation-induced.

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Differentiation-induced changes in Tfam mRNA stability (A) The nt sequence corresponding to the 3′-UTR of mouse Tfam is illustrated. Differentiation-induced changes in Tfam mRNA stability (A) The nt sequence corresponding to the 3′-UTR of mouse Tfam is illustrated. Possible binding sites for RNA-binding proteins, HuR, CuGBP1 and KSRP are underlined. Primers used for 746 and 600 bp Tfam 3′-UTR constructs are highlighted in grey for the forward reaction and black for reverse reaction. (B) Linear regression analysis of Tfam mRNA availability in actinomycin D-treated day 1 compared with day 4 differentiated C2C12 cells (n=3). Slopes of day 1 (−14.45±2.657) and day 4 (−4.475±1.039) decay rates differentiated cells are 3.2-fold different (n=3 experiments). (C) Effect of differentiation on Renilla luciferase activities of 746 and 600 bp Tfam 3′-UTRs. (Day 3–4 compared with day 0–2; n=4, ¶P<0.001; 746 compared with 600 bp Tfam 3′-UTR; n=4, *P<0.001). Values are expressed as means±S.E.M. (D) Total cellular protein levels of HuR, CuGBP1 and KSRP at days 0, 2 and 4 of differentiation. Representative blots of seven experiments are shown. Melania Collu-Marchese et al. Biosci. Rep. 2015;35:e00221 ©2015 by Portland Press Ltd