Fig. 2.

Human and mouse SEC23A and SEC23B complement the yeast Sec23 protein. (A) The coding sequences of the human (h) or mouse (m) SEC23 paralogs (as well as yeast Sec23) were cloned with a 3′ sequence encoding an HA-tag into the pRS416 vector downstream of a ZEO1 promoter. In contrast to WT yeast, temperature-sensitive sec23-1 mutant yeast transfected with a control “empty vector” grow at the permissive (24 °C) but not at the restrictive (37 °C) temperature. Growth of temperature-sensitive sec23-1 mutant yeast expressing h or m SEC23A/SEC23B (or yeast Sec23) was determined at the permissive and restrictive temperatures. Yeast cultures were plated as 10-fold serial dilutions. (B) Immunoblotting for HA (with Pgk1 as loading control) detects relative expression for each construct.