Figure S3.
Meiosis I cohesin dynamics and regulation during meiosis I. (A) Kymographs of anaphase onset in embryos expressing GFP::COH-3 (green) and H2B::mCherry (magenta). Time (s) is relative to anaphase I onset (t = 0). GFP::COH-3 remains high until seconds before anaphase I onset in WT, is reduced before anaphase onset in ify-1(RNAi) embryos, and persists past anaphase I in sep-1(RNAi) embryos. (B) Representative image of GFP::COH-3 (green) and H2B::mCherry (magenta) showing a folded short arm (yellow caret) from different orientations on three bivalents. (B′) Proposed model of GFP::COH-3 at the bivalent short arm adopting a folded configuration based on images from 10 embryos at several stages of meiosis I. (C) In control, GFP::COH-3 midbivalent signal is similar in oocytes (−1) and prometaphase I embryos (labeled 0, white arrows) but is absent from embryos after meiosis I in the uterus (+1, +2). (D) After apc-2 RNAi, GFP::COH-3 levels are retained at the midbivalent in prometaphase I embryos (0) and several arrested embryos in the uterus (+1, +2). (E) GFP::COH-3 midbivalent signal is prematurely reduced after ify-1 RNAi, in prometaphase I embryos (labeled 0) relative to oocytes (−1) and is not observed in older embryos (+1). (F) Quantification of GFP::COH-3 levels relative to H2B from a time series of control or ify-1(RNAi) embryos. Time (s) is relative to anaphase I onset. (G) Quantification of the ratio between the GFP::COH-3 signal at the midbivalent in prometaphase I embryo relative to the −1 oocyte in the same plane. The GFP::COH-3 signal ratio was near 1 in control (N = 10) and apc-2 RNAi (N = 5, P > 0.05) but was significantly reduced (about 50% reduction) after ify-1 RNAi (N = 13, P < 0.05). Scale bars: 5 µm.

Meiosis I cohesin dynamics and regulation during meiosis I. (A) Kymographs of anaphase onset in embryos expressing GFP::COH-3 (green) and H2B::mCherry (magenta). Time (s) is relative to anaphase I onset (t = 0). GFP::COH-3 remains high until seconds before anaphase I onset in WT, is reduced before anaphase onset in ify-1(RNAi) embryos, and persists past anaphase I in sep-1(RNAi) embryos. (B) Representative image of GFP::COH-3 (green) and H2B::mCherry (magenta) showing a folded short arm (yellow caret) from different orientations on three bivalents. (B′) Proposed model of GFP::COH-3 at the bivalent short arm adopting a folded configuration based on images from 10 embryos at several stages of meiosis I. (C) In control, GFP::COH-3 midbivalent signal is similar in oocytes (−1) and prometaphase I embryos (labeled 0, white arrows) but is absent from embryos after meiosis I in the uterus (+1, +2). (D) After apc-2 RNAi, GFP::COH-3 levels are retained at the midbivalent in prometaphase I embryos (0) and several arrested embryos in the uterus (+1, +2). (E) GFP::COH-3 midbivalent signal is prematurely reduced after ify-1 RNAi, in prometaphase I embryos (labeled 0) relative to oocytes (−1) and is not observed in older embryos (+1). (F) Quantification of GFP::COH-3 levels relative to H2B from a time series of control or ify-1(RNAi) embryos. Time (s) is relative to anaphase I onset. (G) Quantification of the ratio between the GFP::COH-3 signal at the midbivalent in prometaphase I embryo relative to the −1 oocyte in the same plane. The GFP::COH-3 signal ratio was near 1 in control (N = 10) and apc-2 RNAi (N = 5, P > 0.05) but was significantly reduced (about 50% reduction) after ify-1 RNAi (N = 13, P < 0.05). Scale bars: 5 µm.

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