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. 2014 Aug 25;15(9):14997–15010. doi: 10.3390/ijms150914997

Figure 4.

Figure 4

p62 promotes the aggresome formation of ataxin-3-80 depended on microtubule. (A) p62 does not affect the self-aggregation of ataxin-3-Q80. H1299 cells were transfected with si-nc or si-p62 siRNA (1# plus 2#) for 12 h, then co-transfected the plasmids of Flag-ataxin-3-Q80 and EGFP or ataxin-3-Q80-EGFP for another 48 h. The cell lysates were subjected to immunoprecipitation assays with anti-GFP antibody. The immunoprecipitants were subjected to immunoblot analysis with the indicated antibodies; (B) HEK293 cells were co-transfected the plasmids of Flag-p62 and ataxin-3-Q80-EGFP for 48 h, then treated with MG-132 (10 µM) or MG-132 (10 µM) combined with nocodazole (5 µg/mL) for another 12 h. Then the cells were fixed and subjected to immunofluorescent assay with anti-Flag antibody (Blue). The scale bar indicates 10 µm; (C) The quantifications of three different distribution patterns of ataxin-3-Q80-EGFP in (B). Three independent experiments were performed and the data are indicated as the means ± S.E.M.