Figure 3.

BDNF selectively enhances Arc label incorporation and expression. Arc protein incorporation and net protein expression are analyzed with autoradiography (A) followed by immunoblot (B) of the same membrane after 2D electrophoresis. Both ³⁵S incorporation and overall Arc protein expression measured on blots, marked with circles, were significantly increased by BDNF treatment. However, BDNF treatment reduced the incorporation of ³⁵S into spots a (0.86), b (0.64), c (0.82), and d (0.91) and increased spots e (1.25) and f (2.74), compared with the control. BDNF did not significantly increase β-tubulin levels overall, and in the experiment represented in B actually decreased β-tubulin by 37% of the control. (C) Summary (range, quartiles, median) of independent experimental results obtained from both autoradiography and immunoblots. The variance of basal Arc protein levels in SNSs was ± 2.1% (SEM) as determined with immunoblots (n = 3) normalized to synaptotagmin levels. On average, BDNF increased Arc ³⁵S incorporation and overall Arc protein levels by 43% ± 4.8% (n = 8, P < 0.01) and 70% ± 9.13% (n = 12, P < 0.01), respectively, relative to paired controls. However, BDNF treatment did not change synaptotagmin (99% ± 1.4%, n = 7, P > 0.05) and β-tubulin (90% ± 7.7%, n = 8, P > 0.05) protein levels compared with controls.