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. 2020 Jun 9;9:e56611. doi: 10.7554/eLife.56611

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© 2020, Srinivasan et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 3. — (A) Exponentially growing wild type (K23889), mrc1Δ (K28092), chl1Δ (K28082), ctf4Δ (K28084), csm3Δ (K28108), tof1Δ (K28091) and ctf18Δ (K28115) expressing endogenous 6C cohesin were synchronised in G1, released into a G2/M arrest and subjected to in vivo crosslinking and mini-chromosome IP. The CM and CD bands are marked. Data shown is representative of two independent biological repeats. (B) Wild type (K24697) mrc1Δ (K28276), chl1Δ (K28175), ctf4Δ (K28275), csm3Δ (K28282), tof1Δ (K28280) and ctf18Δ (K28285) strains that contain genes coding for 6C non cleavable cohesin (2C SMC1 2C SMC3 and GALp-2C SCC1^NC) were arrested in G2 phase, the expression of 2C Scc1^NC was induced by addition of galactose for 45 min. The cultures were released from the G2 arrest and arrested in the subsequent the G1 phase, mini-chromosome IP of the cultures at this stage shows CMs formed in the previous G2 phase being retained in the subsequent G1 in all the strains. The cultures were released from the G1 arrest and allowed to undergo replication and subjected to mini-chromosome IP, the CM and CD bands are marked. A species migrating between the CMs and supercoiled mini-chromosome DNA routinely detected in the ctf18Δ strain is marked with *. We currently do not know the identity of this band. Data shown is representative of two independent biological repeats.

Figure 3—figure supplement 1. — (A) Exponentially growing wild type (K23889), mrc1Δ (K28092), chl1Δ (K28082), ctf4Δ (K28084), csm3Δ (K28108), tof1Δ (K28091) and ctf18Δ (K28115) expressing endogenous 6C cohesin were synchronised in G1, released into a G2/M arrest and subjected to in vivo crosslinking and mini-chromosome IP. The CM and CD bands are marked. Data shown is representative of two independent biological repeats. (B) Wild type (K24697) mrc1Δ (K28276), chl1Δ (K28175), ctf4Δ (K28275), csm3Δ (K28282), tof1Δ (K28280) and ctf18Δ (K28285) strains that contain genes coding for 6C non cleavable cohesin (2C SMC1 2C SMC3 and GALp-2C SCC1^NC) were arrested in G2 phase, the expression of 2C Scc1^NC was induced by addition of galactose for 45 min. The cultures were released from the G2 arrest and arrested in the subsequent the G1 phase, mini-chromosome IP of the cultures at this stage shows CMs formed in the previous G2 phase being retained in the subsequent G1 in all the strains. The cultures were released from the G1 arrest and allowed to undergo replication and subjected to mini-chromosome IP, the CM and CD bands are marked. A species migrating between the CMs and supercoiled mini-chromosome DNA routinely detected in the ctf18Δ strain is marked with *. We currently do not know the identity of this band. Data shown is representative of two independent biological repeats.