Figure 9.

Expression of CLN2 from a heterologous promoter induces bud formation and actin polarization in sda1-2 cells at the restrictive temperature. (A) G0-arrested sda1-2 cells containing GAL1–3XHA-CLN2 were released into fresh media containing glucose or galactose at the permissive or the restrictive temperatures for 8 h and levels of 3XHA-Cln2 were measured by Western blotting with anti-HA antibodies. (B) G0-arrested sda1-2 cells containing GAL1–3XHA-CLN2 were released into fresh media containing glucose or galactose at the restrictive temperature. Pictured are Nomarski images and actin staining in cells incubated for 6 h in glucose-containing media or for 6 and 24 h in galactose-containing media. (C) DNA content measured by fluorescence-activated cell sorter analysis of sda1-2 cells containing GAL1–3XHA-CLN2 in G0 or released into fresh media containing galactose at the permissive or restrictive temperatures for 8 h.