PPA induces accumulation of TH mRNA and TH protein in vitro.

<p>PC12 cells were treated with SCFA: propionic acid (PPA, 1 mM), butyric acid (BA, 1 mM) or valproic acid (VPA, 0.1 mM). At indicated times (48 hrs) total RNA or total cell lysates (0, 24, 48 and 72 hrs) were prepared and subjected to Northern (A) or Western blot analyses (B, PPA and BA). The data on <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0103740#pone-0103740-g002" target="_blank">Fig. 2A</a> are presented as mean ± SEM value relative to the mRNA levels in control, vehicle treated cells. 18S rRNA was used as a house keeping gene for normalization of data. *P<0.05 SCFA vs. vehicle treated. For Western blotting (2B) cells were lysed after indicated incubation times and equal amounts of proteins were subjected to analyses using TH-specific antibody as described in Experimental procedures. Membranes were re-probed with β-actin specific antibody as loading control. The autoradiographs were analyzed by densitometry. Values are presented as the ratio of TH to that of beta-actin as fold difference from control, vehicle- treated cells. N = 6–8 independent samples per group, and the experiment was repeated twice with similar results <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0103740#pone.0103740-Parab1" target="_blank">[63]</a>.