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.
Strating and Martens BMC Cell Biology 2009 10:35 doi:10.1186/1471-2121-10-35