LiCl increases PGC-1α protein expression in VSMCs.

<p>Cells were pretreated with 15 mmol/L LiCl for 30 min and then stimulated with FBS for 24 hr unless otherwise indicated. The mRNA and protein expression levels of PGC-1α were determined by RT-qPCR (A, B) and Western blot (C, D), respectively. PGC-1α protein levels in a cycloheximide chase experiment (E). To determine the half-life of PGC-1α protein, the pixels for each band were measured and normalized so that the number of pixels at <i>t</i> = 0 was 100%. The log₁₀ of the percent of pixels was plotted <i>versus</i> time for each time point, and the <i>t</i>1/2 was calculated from the log of 50%. PGC-1α protein ubiquitination determined by coimmunoprecipitation assay (F). The cellular localization of PGC-1α was shown by immunocytochemistry (G). The red and blue area represents PGC-1α protein and DAPI-stained nuclei. Scale bar indicates 50 µm. CTL: control, cells were treated with 15 mmol/L NaCl. Data are reported as means ± SEM from three independent experiments.