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The magnetic properties of the title complexes were studied by EPR spectroscopy, magnetometry and ab initio calculations., Tetraoxolene radical-bridged lanthanide SMM systems were prepared for the first time by reduction of the respective neutral compounds. Magnetic measurements reveal the profound influence of the radical center on magnetic behavior. Strong magnetic couplings are revealed in the radical species, which switch on SMM behavior under zero applied field for DyIII and TbIII compounds. HFEPR spectra unravel the contributions of the magnetic coupling and the magnetic anisotropy. For GdIII this results in much more accurate magnetic coupling parameters with respect to bulk magnetic measurements.

The yield of soluble chromatin is increased in the range of tested temperatures (25 °C -65 °C). Two 20-μm sections from mouse liver FFPE tissues were processed by Chrom-EX PE at indicated temperatures. To accurately measure the impacts of incubation temperature on chromatin yield, we isolated the chromatin by sonication but no MNase treatment after tissue-level cross-linking reversal. DNAs were purified from soluble fraction and insoluble pellet fraction and were quantified using Qubit dsDNA High Sensitivity assay. The percentage of soluble chromatin was calculated from two independent experiments. (PDF 50 kb)

DNA profiles for chromatin inputs from 4 liver FFPE tissues relative to Fig. 2a. Two 20-μm sections from mouse liver FFPE tissues were processed by Chrom-EX PE at 65 °C condition. 2.5% input were decross-linked and purified by MinElute PCR Purification Kit and eluted in 16 μl TE and 2 μl DNAs were analyzed by the Fragment Analyzer. (PDF 31 kb)

The genomic location of primer pair along with the peak profiles of H3K4me3 and H3K27me3 marks are visualized for ChIP-seq data generated from frozen and FFPE liver tissues in the Integrative Genomics Viewer. The arrow below the RefGene track indicates the location of primer pair used in the study. The peak profiles indicate the antibodies for H3K4me3 and H3K27me3 marks are specific. (PDF 49 kb)

DNA profiles in RNAP II, H3K27Ac, H3K4me3 and H3K27me3 ChIP-seq libraries from mouse liver FFPE and frozen tissues. Lane 1, 2: RNAP II libraries from FFPE and frozen tissues; Lane 3â 5: H3K27Ac, H3K4me3 and H3K27me3 libraries from FFPE tissues; Lane 6â 8: H3K27Ac, H3K4me3 and H3K27me3 libraries from frozen tissues. (PDF 38 kb)

ChIP enrichment analyzed by qPCR. The ChIP DNA was analyzed by qPCR in a region of constitutively active chromatin region (GAPDH-TSS, H3K4me3-positive), a developmentally repressed region (MYT1-TSS, H3K27me3-positive), and an intergenic region (C19, H3K4me3- and H3K27me3-negative) from HeLa cells. Enrichment in the tested loci is shown as the percentage of input. (PDF 87Â kb)

DNA profiles in H3K4me3 and H3K27me3 ChIP-seq libraries generated through different purification reagents. After library amplification with 12 cycles of PCR, DNAs were analyzed by the Fragment Analyzer. Lane 1 (EDL St), the library from 1 ng of purified ChIP DNA from aliquot A; Lane 2 (1 ng QmM), the library from stored DNA in MaxyClear tube after purification by MinElute PCR Purification Kit from aliquot B; Lane 3 (1 ng QmP), the library from stored DNA in Premium Tube after purification by MinElute PCR Purification Kit from aliquot B; Lane 4 (Input), the library from 1 ng of purified input DNA from the aliquot A. Left panel is the profile of H3K4me3 libraries, and right panel is the profile of H3K27me3 libraries. (PDF 123 kb)

DNA purification reagents vary in their ability to recover low amounts of DNA. a Recovered DNA amount by different DNA purification reagents from de-crosslinked chromatin. De-crosslinked chromatin estimated to include 5, 10, 50 ng range of DNA in ChIP elution buffer was purified following the manufacturer’s instructions as described in Fig. 1a. The data were generated from triplicate DNA samples derived from three independent preparations. b Recovery rates of nanogram-range DNA by different DNA purification reagents. DNAs were adjusted to final 1 ng, 5 ng, 10 ng and 50 ng in 100 μL ChIP elution buffer and were purified by purification reagents following the manufacturer’s instructions. Percent recovery was calculated from DNA amounts before and after purification. The data were generated from triplicate DNA samples derived from three independent preparations. (PDF 105 kb)

Measuring global ChIP enrichment by FRiP (fraction of reads in peaks). The score of FRiP (fraction of reads in peaks) was calculated by the previously published method [3]. FRiP scores for H3K4me3 and H3K27me3 ChIP-seq are visualized from the ENCODE reference (ENB and ENw) and the corresponding datasets obtained either with standard protocol for the lab (St) or by using different purification reagents as identified in Fig. 1. (PDF 94Â kb)

ChIP-seq data generated with different purification reagents are highly correlated with the corresponding ENCODE datasets. Scatter plots showing the correlation between the ENCODE reference ChIP-seq data (ENB and ENw) and the corresponding datasets obtained either with standard protocol for the lab (St) or by using different purification reagents as identified in Fig. 1. The genome was divided into 5Â kb bins for H3K4me3 and 100Â kb bins for H3K27me3, and the number of mapped reads in the individual bins was calculated. r, Pearson correlation coefficient. P value in all correlation analysis was 0.001. A and B panels show H3K4me3 and H3K27me3 ChIP-seq data, respectively. (PDF 248Â kb)

Overlapping percentage of peaks relative to ENCODE data. (PDF 91Â kb)