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. 2014 May 29;3(6):542–552. doi: 10.1242/bio.20148367

Fig. 8. Determination of Cat1 distribution by its modification at the N-terminal lysine residues.

Fig. 8.

(A) Schematic representation of substitutions of lysine residues by arginine in the N-terminal of Cat1. Double-head arrows show the extent of the respective Cat1 mutants including the substitutions of lysine residues. TM denotes a transmembrane domain. (B) JUp1209 cells carrying pREP273-Cat1-WT or the mutants fused with EGFP as indicates were grown in EMM + uracil. (C) L968 (non-tag), AN0541 (Cat1-WT), AN0542(Cat1-a), and AN0543(Cat1-a-b-c) were grown in EMM. Protein extracts were prepared from membrane rich fractions, and proteins were probed with the GFP antibody. (D) JUp1209 carrying pREP273-Cat1-WT or -a-EGFP was grown in EMM + uracil (0 h) and then treated with 30 µg/ml cycloheximide for 1 hour (CHX). (E) JUp1209 carrying pREP273-Cat1-WT or the mutants as indicates were grown in EMM + uracil. (F) JUp1209 carrying pREP273-Cat1-WT- or -b-c-EGFP was grown in EMM + uracil (+N), washed, and incubated in EMM-N for 1 hour (−N). (B,D–F) The GFP images are shown inverted for clarity. Scale bars: 10 µm.