Clp1 requires Mid1 association for CR localization. (A) Live-cell GFP and light images of mid1Δ431–481-GFP cells. (B) Denatured protein extracts were prepared from mid1-GFP and mid1Δ431–481-GFP cells and analyzed by immunoblotting with GFP and dm1a (α-tubulin) antibodies. (C) Live-cell imaging of GFP-Myo2, Rlc1-GFP, and Plo1-GFP in mid1Δ431–481 cells. Arrows indicate rings. Asterisk indicates constricting ring. (D) Live-cell imaging of clp1-GFP mid1Δ431–481 cells incubated at 32°C and nda3-KM311 clp1-GFP mid1Δ431–481 cells after incubation at 18°C for 7 h. Bars, 5 μm. (E) nda3-KM311 clp1-GFP (mid1+), nda3-KM311 clp1-GFP mid1Δ431–481, and nda3-KM311 clp1Δ strains were arrested by incubation at 18°C for 7 h, and then samples were collected and denatured protein extracts were analyzed by immunoblotting with Cdc15 and dm1a antibodies.
