Extended Data Fig. 1: Peroxisomal protein import reactions in Xenopus egg extract and purity of recombinant proteins used in this study.
From: Import mechanism of peroxisomal proteins with an N-terminal signal sequence
a, Xenopus egg extract was depleted or not (mock) of endogenous PEX5 using beads conjugated to the PEX5-binding domain from PEX14, then supplemented with buffer or the indicated purified recombinant PEX5 variants (described in Fig. 1b). Import was assessed by imaging the indicated fluorescent cargo on a spinning-disk confocal microscope. The cargo, shown on the left, consisted of GFP fused to the SKL peroxisome targeting signal (PTS1) or to the signal from the indicated Xenopus PTS2 peroxisomal matrix enzymes (PTS2). Repeated rounds of cargo import into peroxisomes produce bright puncta. Scale bars equal 5 µm. b, The indicated recombinant proteins were produced and purified as described in the Methods, then resolved by SDS–PAGE and stained by Coomassie Blue. Molecular weights (in kD) are marked on the left; images of the unprocessed gels are provided.
