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. 2010 Oct 1;21(19):3330–3339. doi: 10.1091/mbc.E10-04-0274

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© 2010 by The American Society for Cell Biology

This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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Figure 5. — Excess Ac45 affects the processing of newly synthesized POMC in transgenic melanotrope cells. (A) Neurointermediate lobes (NILs) from wild-type (wt) and Ac45-transgenic (tg) animals were pulse labeled for 30 min and subsequently chased for the indicated time periods. 5% of the newly synthesized proteins extracted from the lobes and 20% of the proteins secreted into the incubation medium were directly resolved by 12.5% SDS-PAGE and visualized by autoradiography. The experiments were performed in triplicate, and a representative example is shown. (B) The processing rate of 37-kDa POMC to 18-kDa POMC and the secretion of 18-kDa POMC were quantified by densitometric scanning using a phosphoimager. For each chase period, the ratio of 18-kDa POMC to 37-kDa POMC was calculated for wild-type (wt, white bars) and transgenic (tg, black bars) NILs. The amount of secreted 18-kDa POMC was calculated relative to the levels of newly synthesized actin and is presented in AU. Note that the newly synthesized Ac45-transgene product comigrated with mature PC2 (Jansen et al., 2008). Shown are the means ± SEM (n = 3). Significant differences are indicated by *(p < 0.05) or **(p < 0.01).