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. 2004 Mar;24(6):2324–2331. doi: 10.1128/MCB.24.6.2324-2331.2004

FIG. 3.

FIG. 3.

Decreased expression of cellular ubiquitin contents in Δlub1 cells. (A) Intracellular expression of monoubiquitin. Wild-type (WT), ubi4-H45, and Δlub1 cells harboring the expression vector alone or the vector for Lub1, Ubi4, or FLAG-ubiquitin were cultured for 18 h. The whole-cell lysates were analyzed by immunoblotting with an antiubiquitin antibody to determine the monoubiquitin expression level. Positions of monoubiquitin (Ub) and FLAG-tagged ubiquitin (asterisk) are indicated. WB, Western blotting. (B) Northern blot analysis of ubi4+ mRNA. The wild-type and Δlub1 cells were cultured at 27 or 36°C, respectively. The total RNA was isolated and analyzed by Northern blotting with DIG-labeled ubi4+ cRNA. Equality of the loading mRNA was confirmed with leu1+ cRNA. (C) Immunoblot analysis of multiubiquitin. The wild-type and Δlub1 cells harboring the expression vector for Lub1 (full length) or Lub1 (230-713) were cultured for 18 h. The determination of multiubiquitin expression level is as described for panel A.