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Figure 4 | BMC Cell Biology

Figure 4

From: Incomplete posttranslational prohormone modifications in hyperactive neuroendocrine cells

Figure 4

Glycosylation of POMC in cycloheximide-treated melanotrope cells from black-adapted Xenopus. A, Neurointermediate lobes (NILs) from black-adapted Xenopus were pre-incubated, pulse labelled with [35S]methionine/cysteine for 30 minutes and chased for three hours in the absence (control) or presence of cycloheximide (Cyclohex.; drug concentrations indicated above the lanes). Since for the samples of the cycloheximide-treated NILs the intensities of the autoradiographic signals were relatively low, a larger part of the cycloheximide-treated NIL lysates than of the untreated NIL lysates was loaded (indicated below the lanes). The slightly reduced mobility of 37 K POMC in the sample treated with 1 μg/ml cycloheximide was due to a gel problem. B, NILs from black-adapted Xenopus were pre-incubated, pulsed and chased as in panel A in the absence or presence of the indicated concentration of cycloheximide. Aliquots (10%) of the lysates were control-treated (C) or deglycosylated with Peptidyl N-glycosidase F (PNGaseF; F) or Endoglycosidase H (EndoH; H), and analysed by SDS-PAGE and autoradiography. To compensate for the reduced labelling in the presence of cycloheximide, the right panel was exposed twice as long as the left panel.

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