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Fig. 4 | Epigenetics & Chromatin

Fig. 4

From: Inheritance of the CENP-A chromatin domain is spatially and temporally constrained at human centromeres

Fig. 4

The CENP-A domain is inherited at the same overall location on alpha satellite DNA. a Representative chromatin fiber showing CENP-A immunostaining (green) and FISH with an alpha satellite probe to DXZ1 (red). The distance between the edges of DXZ1 and the edge of the CENPA domain was measured for each fiber. b Calculation of the percentage of DXZ1 between its edge and the edge of the CENP-A domain. For this analysis, the orientation of the alpha satellite array (short arm vs long arm of HSAX) was not known. The direction of each fiber was randomly assigned such that half of the fibers were oriented with CENP-A domains closer to the arbitrary left end of the DXZ1 FISH and half were oriented with CENP-A closer to the arbitrarily assigned right edge of the DXZ1 fluorescent signal. c Analysis of CENP-A domain skewness, by non-randomly assigning the shortest distance between the DXZ1 edge and the leading edge of the CENP-A immunostaining as orientation toward the HSAX short arm. This assumption was made based on previous studies suggesting that centromere proteins are biased toward the short arm side of DXZ1 [11, 52]. d Representative chromatin fiber IF-FISH image showing the position of CENP-A immunostaining (green) relative to the proximal short arm landmark BAC RP13-971O21 (red) located adjacent to DXZ1, the alpha satellite array on the human X chromosome. Scale bar is 5 μm. e Measurement of the percentage of DXZ1 between the short arm edge of the array and the edge of the CENP-A domain in two different cell lines. f BAC RP13-971O21 (red line) was used as a landmark representing the short arm side of the DXZ1 array. Genomic distances representing the placement of the left edge of the CENP-A domain relative to the short arm edge of DXZ1 (blue line) on individual chromatin fibers (green ticks) are shown as custom tracks in the UCSC genome browser illustrating the non-random positioning of the CENP-A domain on the short arm side of the DXZ1 array (blue line) for two different cell lines. The average starting edge of the CENP-A domain based on the distance from the short arm edge of DXZ1 is shown as a single black tick under the individual fiber data

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