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Fig. 3. Vps30 and Vps38 protein functions are required for stability of the Golgi-localized Kex2 protease. Wild-type (SEY6210), {Delta}vps30 (JCY300), {Delta}vps38 (PBY58) and {Delta}vps35 (EMY18) cells carrying plasmid pPB1 (KEX2-HA) were grown to early log phase and labeled with [35S] methionine/cysteine at 26°C for 20 minutes followed by the addition of excess non-labeled amino acids (chase). Samples were removed at 0 and 90 minutes after the addition of chase. Kex2-HA (upper panel) was immunoprecipitated from cell lysates using HA-specific antibodies. The same cell lysates were then re-immunoprecipitated with G6PDH-specific antibodies (lower pannel). Kex2-HA and G6PDH were resolved by SDS-PAGE and visualized by autoradiography.