Fig. 4.

Thyroid hormone is a positive regulator of SOD2 transcription.(A) ChIP seq analysis of T3-target genes performed in C2C12 cells showed that the SOD2 gene is a direct target of T3. The mammalian conservation, the gene structure and position from Genome Browser and the ChIP Seq results (black bars) are indicated for the SOD2 gene. (B) Chromatin immunoprecipitation assay was performed in C2C12 cells. Immunoprecipitation of the chromatin using the anti-TRα antibody revealed that the SOD2 gene is a direct target of T3. (C, D) Protein levels of SOD2 were measured by Western blot analysis in pTRE-D2 cells treated or not with doxycycline and in cells treated with TH for 10 h and 24 h. (E) Immunofluorescence analysis of SOD2 expression in pTRE-D2 cells treated or not with doxycycline. (F, G) Real time PCR analysis and Western blot analysis of SOD2 expression in gastrocnemius (GC) muscles of hyper- and hypothyroid mice (see Methods). (H) SOD2 expression was analyzed in muscle fibers isolated from control (euthyroid) and hyperthyroid mice. (I) mitoSOX analysis in the same muscle fibers as in H was assessed by immunofluorescence. The results are shown as means ± SD from at least 3 separate experiments. *P < 0.05, **P < 0.01.