Fig. 7
From: Accumulation of the PX domain mutant Frank-ter Haar syndrome protein Tks4 in aggresomes
The proteasome inhibitor MG132 increases the level of aggresome formation by the Tks4R43W mutant. 7a, COS7 cells were transiently transfected with a V5-Tks4 and V5-Tks4R43W construct. After 18 h, the cells were treated with 5 μM MG132 for 20 h. The cells were fixed and stained for Tks4 (with anti-V5 antibody, red) and α-tubulin (green). Cell nuclei were visualized by DAPI staining. 7b, The percentage of cells with aggresomes was quantified by counting 300 cells/sample by using the ImageJ software. 7c, COS7 cells were transiently transfected with V5-Tks4 and V5-Tks4R43W construct. After 18 h the cells were treated with 5 μM MG132 for 20 h or left untreated. Cells were then lysed, and the lysates were separated into detergent-soluble (S) and insoluble pellet fractions (P) by centrifugation and analyzed by Western blotting for Tks4 (with anti-V5 antibody), Cbl, caveolin-1 and HDAC6, respectively. 7d, The Tks4 protein levels were determined by densitometry. Error bars represent the standard deviation from three independent experiments. Asterisks indicate the level of significance (one asterisk, P < 0.05; two asterisks, P < 0.01; three asterisks, P < 0.001)