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We also analyzed the previously published transcription factor ChIP-seq datasets for binding at centromeric sites (Gerstein et al., 2010).
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Comparison of input signal at centromeric sites in native and formaldehyde-crosslinked samples.
(A, B, C ) Normalized fragment counts in input (A ), cenH3 ChIP (B ) and insoluble chromatin (C ) samples at centromeric sites.
Embryonic cells contain both cenH3 and transcription factors, thus complicating the analysis of the chromatin landscape at centromeric sites.
These results confirm the findings in other organisms that cenH3 nucleosomes at centromeric sites in C. elegans wrap less DNA than that wrapped by canonical nucleosomes.
We found that all tested transcription factors profiled in larval instar 2 or later stages, when few somatic cells are still dividing, are enriched at centromeric sites.
In total adult samples, which contain nuclei from dividing germline and embryonic cells, we found a depletion of signal at centromeric sites and well-positioned nucleosomes flanking the sites, reminiscent of the chromatin landscape in embryos.
At some centromeric sites in transformed/immortalized cell lines, H3K4me2 and heterochromatic modifications appeared to coincide.
Relative occupancy at a chromosome arm site (c194) or at a centromeric site (CENV) was determined relative to a low binding region (c281).
Relative occupancy at a centromeric site (CEN5) relative to a low binding region (HMR) was determined.
Spo13-3V5 localization was determined by ChIP 7 hr after transfer into sporulation medium when cells were arrested in metaphase I. Relative occupancy at a centromeric site (CENV) relative to a low binding region (HMR) was determined.
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