Figure 3.

RAB6A Validation as a Substrate for In Vitro Prenylation by Other Cell Lines

Protein expression (human REP1 and β-actin) and biotin incorporation were detected in prenylation reaction products following cell transduction, SDS-PAGE, and western blot analysis (two replicates in one experiment). HT-1080 cells (A) and ARPE-19 cells (B) were transduced with rAAV2/2-REP1 (MOI 1,000; 10,000; and 30,000 gc/cell), and prenylation reactions were prepared with 20 μg and 10 μg of total protein, respectively. Positive controls (+ rREP1) were prepared using untransduced cell lysate spiked with a recombinant fish REP1 protein (25 nM for HT-1080; 11 nM for ARPE-19).