FIG 8.

Isolation of p53 amyloids from yeast cell lysate. (A) Amyloid-containing fractions were isolated from yeast cell lysates by sedimentation in SDS-containing lysis buffer. The p53 protein was detected in the total lysate, the SDS-soluble supernatant, and the SDS-insoluble pellet fraction by immunoblotting with anti-p53 antibody. A prestained molecular mass ladder was loaded in the middle lane. The blot was developed with the colorimetric substrate TMB (Genei, India). The blot was developed with anti-GPD to assess GPD levels in fractions with and without seeds as a loading control. (B) Lysates of yeast cells with/without seeds were analyzed by SDD-AGE and Western blotting. The p53 expression was detected by immunoblotting with anti-p53 antibody. Cell lysate from plus-seed cells showing higher-order SDS-resistant aggregates are marked in the gel.