Figure 5: Time course of ANL labelling of neuronal proteins in adult flies.
From: Cell-selective labelling of proteomes in Drosophila melanogaster
(a) Zero to three days old elavC155-Gal4;;UAS-mMetRSL274G-EGFP flies were placed either onto ONM with 4 mM ANL or without ANL. After 24 and 48 h, heads from adult Drosophila flies were collected and ANL incorporation was addressed by BONCAT. Representative immunoblots showing the extent of ANL-labelled proteins and the neuronal marker protein synapsin after NeutrAvidin purification. The signal intensities for both overall newly synthesized proteins and synapsin increases from 24 to 48 h of ANL incorporation periods. (b) The assessment of protein-incorporated ANL into proteins and into synapsin, which is transmitted from larval neurons throughout the pupal phase into adult brain proteins, is rather high compared with flies that were on ANL-containing ONM through all developmental stages. Depicted immunoblots show input (I, before NeutrAvidin purification) and eluted fractions (E, enriched ANL-labelled proteins after NeutrAvidin purification) of samples.+ represents 4 mM ANL and − represents w/o ANL.
