Figure 6

Polysomal distribution of DZIP1-GFP. (A) Lysates were prepared from HeLa cells treated with cycloheximide and were separated in a 10–50% sucrose gradient. Distribution of DZIP1-GFP was examined by western blotting. (B) HeLa cells were treated with puromycin, and extracts were prepared and processed as in (A). (C) Quantitation of the distribution of DZIP1-GFP after cycloheximide and puromycin treatment. Western blots shown in (A) and (B) were used for quantitation. (D) Protein extracts of Hela cells transfected with pDZIP1-GFP, pGFP alone (control sample), siDZIP1 or siNC1 were analyzed by western blotting with antibodies against SNX2, GAPDH1 and GFP. Quantitation by integrated density of SNX2, DZIP1-GFP (α-GFP ~120 kDa) or GAPDH1 is shown in the table above.