Combined modulations of membrane anchoring and N-terminal clustering strongly affect Cdr2 node assembly. (A) Localization of mEGFP-tagged Cdr2, Cdr2^bsc-3D, and Cdr2^bsc-3A in wild-type and mid1^Δ400–450 cells or when Cdr2 also carries the E177A kinase-dead mutation (KD). (B) Number of nodes in the medial focal plane relative to cell length in similar cells as in A. Error bars show SDs. (C) Differential interference contrast images of mid1^nsm or cdr2^bsc-3A mid1^nsm cells (left) and asymmetric positioning of the division plane measured as a ratio between the distance between septum and closest cell tip over total cell length in mid1^nsm (n = 48), cdr2^bsc-3A (n = 53), cdr2^bsc-3A mid1^nsm (n = 53), and pom1Δ cells (n = 64) from one experiment out of three repeats (right). Asterisks show cells with an asymmetrically positioned septum. Horizontal bars are means. t tests: *, P < 5 × 10⁻²; ***, P < 10⁻⁵. Bars, 5 µm.