Ags1 localization in the division site depends on the CAR formation, but not on the SIN pathway. (A) cdc12-112 ags1⁺-GFP cells were shifted to 32°C for 4 h. (B) cdc15-140 ags1⁺-RFP rlc1⁺-GFP cells were shifted to 36°C for 30 min and imaged by time lapse. (C) cdc11-119 ags1⁺-RFP rlc1⁺-GFP and (D) cdc11-119 ags1⁺-GFP hht1⁺-RFP cells were shifted to 36°C for 1 h. (E) cdc15-140 ags1⁺-RFP rlc1⁺-GFP cells were shifted to 32°C for 30 min. Square: magnified area. Arrow: Ags1 localization in the medial region. Arrowhead: unstable CAR appearance in the middle of the cell. Elapsed time is shown in minutes. Bars, 5 µm.