Figure 2.
The chc1-box mutant is not temperature-sensitive for growth. (A) SL82 (chc1Δ + pAP4[CEN, CHC1]), SL5177 (chc1Δ + pJRC2[CEN, chc1-1]), SL5171 (chc1Δ + pJRC3[CEN, chc1-2]), SL5181 (chc1Δ + pJRC4[CEN, chc1-3]), SL5178 (chc1Δ + pJRC5[CEN, chc1-4]), and SL5634 (chc1Δ + pJRC19[CEN, chc1-box]) cells were diluted to 107 cells/ml, and fourfold serial dilutions were spotted onto plates and grown at the indicated temperatures for 3 d. (B) Immunoblots of extracts from chc1Δ yeast (SL12) transformed with pUN30 (CEN, empty), pAP4 (CEN, CHC1), pJRC2 (CEN, chc1-1), pJRC3 (CEN, chc1-2), pJRC4 (CEN, chc1-3), pJRC5 (CEN, chc1-4), or pJRC19 (CEN, chc1-box) were detected with anti-Chc1 antibodies. Cells were grown continuously at 25°C, and then one-half of each sample was removed and incubated for 90 min at 37°C before harvesting. To verify equal protein loading, blots also were probed with antibodies to tubulin.