Figure 5. Loss of Fic1 impairs CR disassembly and leads to persistence of division site factors.

(A) Fixed-cell DIC and DAPI/methyl blue images of asynchronous and G2-arrested cells of the indicated genotypes. Arrowheads indicate cells that are still joined following ingression. (B) Quantification of (A), with four trials per genotype and n>300 for each trial. Percentages are presented as mean ± SEM. (C) Live-cell GFP movies of rlc1-GFP sid4-GFP and fic1Δ rlc1-GFP sid4-GFP cells. Images were acquired every 2 min, and representative images are given for 10 min intervals. (D) Quantification of times from spindle pole body (SPB) separation to the completion of CR constriction in (C). n>20 for each genotype. Data are presented in box-and-whisker plots showing the median (line in the box), 25^th–75^th percentiles (box), and 5^th–95^th percentiles (whiskers) for each genotype. (E) Quantification of times from septum closure to disappearance of the CR at the division site for GFP-cps1 rlc1-mCherry₃ and fic1Δ GFP-cps1 rlc1-mCherry₃ cells. n>30 for each genotype. Data are presented in box-and-whisker plots showing the median (line in the box), 25^th–75^th percentiles (box), and 5^th–95^th percentiles (whiskers) for each genotype. (F) Live-cell GFP (colored green) and mCherry (mCh) (colored magenta) movies of GFP-cps1 rlc1-mCherry₃ and fic1Δ GFP-cps1 rlc1-mCherry₃ cells, with time intervals indicated and GFP/mCherry images merged. White arrows in GFP images mark the septa's leading edges. The time point with only one arrow drawn marks septum closure. In the mCh images, “C” marks the point of CR closure, and arrowheads denote CR remnants persisting after this point. (G) Fixed-cell images of actin stained with Alexa Fluor 488 Phalloidin. Single z planes as well as maximum projections of multiple z planes are given. Red arrows indicate division planes, whereas yellow arrows indicate unusual actin masses lining the division plane (Bars = 5 µm).