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Fig. 6 | Epigenetics & Chromatin

Fig. 6

From: 27nt-RNAs guide histone variant deposition via ‘RNA-induced DNA replication interference’ and thus transmit parental genome partitioning in Stylonychia

Fig. 6

The nanochromosome copy number in vegetative macronuclei is an indicator for the existence of ‘RNA-induced DNA replication interference’ following S-phase treatments. a Replication in vegetative macronuclei (M) takes place in replication bands comprising of a forward zone (fz) with high DNA concentration and a rear zone (rz) with low DNA concentrations. The fz determines the direction of migration (arrow). Replication bands do never appear in micronuclei (m). b The motility of chromatin in macronuclei is strictly constrained, as revealed through pulse-chase-pulse experiments, where newly replicated DNA was 30 min pulse-labelled with 5-iodo-2′-deoxyuridine (red signals), followed by a 2 h chase and then a 30 min pulse with 5-chloro-2′-deoxyuridine (yellow signals): The signals from the first pulse remain within a restricted area even after prolonged observation periods/chases [20]. The arrow indicates the migration direction. a, b Top-Pro-3 was used for DNA counterstaining, false colours were used for illustration. c The chart illustrates the effects of mock/bait RNA treatment (microinjection) on the copy numbers of targeted nanochromosomes at 3 different time periods (t1–3). The chart combines the data from all (target) experiments. The mock median was used as reference (onefold change) for the calculations of relative fold changes. Data was obtained from 48 independent qPCR reactions in triplicate (mock RNA) or 64 independent qPCR reactions in triplicate (bait RNA), whereby three different reporter amplicons were evenly distributed for bait RNA PCR reactions

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