Figure 2.

Immunoblot analysis with anti-Urm1p antibodies. Exponential cultures of haploid strains SY3358 (WT), SY3806 (urm1Δ) and SY3805 (uba4Δ), were lysed in the presence of 20 mM NEM (lanes 1-3) or in the absence of NEM (lanes 4 - 6). Lysates were analyzed by SDS-PAGE and immunoblot analysis by using affinity-purified polyclonal anti-Urm1p antibodies and monoclonal antibodies to Dpm1p (to confirm equal protein loading). The band corresponding to unconjugated Urm1p is indicated. A half-open square bracket designates high molecular mass Urm1p modified species, whereas an asterisk denotes the position of a 32-kDa Urm1p-modified species.