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Image Search Results
Journal: Nature Structural & Molecular Biology
Article Title: Loss of H3K9 trimethylation alters chromosome compaction and transcription factor retention during mitosis
doi: 10.1038/s41594-023-00943-7
Figure Lengend Snippet: a , Volcano plots as in Fig. , highlighting pluripotency-associated factors that are enriched (red) or not significantly changed (black) on WT (left plot) or Suv39h dn (right plot) ESC mitotic chromosomes. b , Esrrb immunolabeling (red) of WT and Suv39h dn flow-sorted chromosomes 19 (left panel) and X (right panel), where DAPI counterstain is shown in light gray. Scale bar, 5 μm. Esrrb mean intensities were measured across individual chromosomes; the mean ± s.d. is shown ( n = minimum 50 chromosomes analyzed over 3 independent experiments). P values of statistically significant changes, measured by unpaired, two-tailed Student’s t -tests, are indicated. c , Esrrb ChIP–qPCR analysis in WT versus Suv39h dn mitotic and asynchronous ESCs. Enrichment (immunoprecipitated as a percentage of input (% IP)) was measured at Esrrb bookmarked sites ( Capn2 , Esrrb , Jam2-s1 , Jam2-s2 , Tbx3 and Tet2 ), Esrrb lost sites (bound only in interphase ; Mgat3 and Twistnb ) or control sites that do not bind Esrrb ( Esrrb-3 ′ and Actb ). The mean + s.d. results are shown. For interphase cells n = 3 biological replicates, for mitotic cells n = 4 biological replicates (except Capn2 , Esrrb , Rex1 and Jam2-s1 , where n = 5). d , Live-cell imaging of Esrrb–tdTomato mouse ESCs pretreated with DMSO (upper panel) or 100 nM of chaetocin (lower panel) cultured with SiR-DNA (red). Arrows show Esrrb localization to mitotic chromatin. Scale bar, 5 μm. Esrrb–tdTomato mean intensities on mitotic DNA (gated based on SiR-DNA signal) and in interphase nuclei were quantified for each condition; the mean ± s.d. is shown. For mitotic chromosomes: n = 25 (DMSO) or n = 35 (chaetocin) cells analyzed; for interphase nuclei: n = 46 cells analyzed for both DMSO and chaetocin treatments, representing 3 independent experiments. b – d , P values of statistically significant changes, measured by unpaired, two-tailed Student’s t -tests, are indicated. and precise n numbers are provided.
Article Snippet:
Techniques: Immunolabeling, Two Tailed Test, ChIP-qPCR, Immunoprecipitation, Control, Live Cell Imaging, Cell Culture