Fig. 2.

Histone deacetylase 4 (HDAC4) increases Gadd45a mRNA in mouse skeletal muscle. A: bilateral TA muscles of C57BL/6 mice were harvested at the indicated times after unilateral hindlimb denervation, and Gadd45a and HDAC4 mRNA levels were quantified by qPCR. At each time point, levels in denervated muscles were normalized to levels in contralateral innervated muscles, which were set at 1. B: in C57BL/6 mice, one TA muscle was transfected with 20 μg p-HDAC4-FLAG, and the contralateral TA (“Control”) was transfected with 20 μg empty plasmid (pcDNA3). At the indicated times after transfection, bilateral TAs were harvested for SDS-PAGE and immunoblot analysis of HDAC4-FLAG expression (top) and qPCR analysis of Gadd45a mRNA expression (bottom). In the immunoblot analysis, actin served as a loading control. At each time point, Gadd45a mRNA levels in the presence of HDAC4 were normalized to levels in the absence of HDAC4. C: in C57BL/6 mice, one TA muscle was transfected with 20 μg p-Gadd45a-FLAG, and the contralateral TA (Control) was transfected with 20 μg empty plasmid (pcDNA3). At the indicated times after transfection, bilateral TAs were harvested for immunoblot analysis of Gadd45a-FLAG expression (top) and qPCR analysis of HDAC4 mRNA expression (bottom). A–C: data are means ± SE from 4–6 mice/time point. Some error bars are too small to see. *P ≤ 0.05.