Figure 5.

Localization of Arl1p on trans-Golgi is nucleotide dependent. Wild-type Arl1, Arl1Q72L, or Arl1T32N from integrated plasmid YIplac128 under the control of the ADH promoter was transformed into arl1 (YPH250dl1) cells expressing GFP-Sft2p from pRS314 vector. Logarithmically growing cells were fixed and spheroplasts were prepared for reaction with affinity-purified anti-Arl1p antibody and monoclonal anti-GFP antibody. Indirect immunofluorescence staining was done as described in Materials and Methods. Observations were replicated in at least three experiments.