Decreased cellular levels of Vps45. (A) Mutation Thr238Asn in yeast Vps45 (equivalent to Thr224Asn in the human protein) results in lower cellular levels of Vps45 and Tlg2. Western blot analysis was used to assess cellular levels of Tlg2 in cells containing (+) or lacking (−) endogenous VPS45. In addition, levels of HA-tagged versions of either wild-type (WT) Vps45 or Vps45T238N (T/N) and Tlg2 were assessed in cells lacking endogenous VPS45 but harboring plasmids pCOG070 (encoding WT Vps45) or pMC007 (encoding Vps45T238N). Levels of Pgk1 were assessed as a loading control in all cases. A representative blot of 3 independent experiments is shown. (B) Western blot of the Vps45 protein is shown. Vps45 was detected by rabbit polyclonal anti-VPS45 antiserum and anti-rabbit IgG horseradish peroxidase followed by an enhanced chemiluminescence reaction. Actin, as a housekeeping gene, is shown at the bottom. Shown are lymphoblasts from 2 patients and from a normal control. MW, molecular weight; HA, human influenza hemagglutinin epitope tag.