(A and B) Loss of essential inner kinetochore component Ame1CENP-U in cycling and prophase I cells lacking IML3 and MCM21. Prophase I-arrested (A) or cycling (B) wild-type, iml3Δ, and mcm21Δ cells carrying AME1–6HA, together with untagged control, were subjected to anti-HA ChIP-qPCR. Error bars represent SE (n = 3 or 4 biological replicates, cycling and prophase I-arrested cells, respectively).
(C–F) Ame1-mNeonGreen imaging in cycling (C and D) and pre-S phase-arrested (E and F) wild-type and mcm21Δ cells. Signal quantification (C and E) and representative images (D and F) are shown. In (C) and (E), whiskers represent 1.5 IQR, the middle line is median, and the box encompasses the two middle quartiles of the data. ****p < 10−10, **p < 10−4; t test. n > 13 (C) or n > 17 cells (E).
(G) Wild-type and mcm21Δ cells expressing Ame1-mNeonGreen were allowed to synchronously enter pre-meiotic S-phase through induction of IME1 and IME4, followed by live-cell imaging.
See also Figure S7.