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. Author manuscript; available in PMC: 2016 Aug 31.
Published in final edited form as: Cell Rep. 2016 Aug 18;16(9):2308–2316. doi: 10.1016/j.celrep.2016.07.079

Figure 1. CENP-A is reduced in genomic regions lost during nematode programmed DNA elimination.

Figure 1

A and B. CENP-A is reduced in genomic regions that that remain at the metaphase plate and will be lost during Ascaris DNA elimination. Immunohistochemical (IHC) staining of CENP-A on a 4-cell Ascaris embryo with two cells undergoing DNA elimination mitoses (anaphase) indicates the DNA that wiil be lost (red arrows) has much less CENP-A than the DNA that will be segregated and retained. The staining observed is the same using two independently prepared antibodies (Ab1 and Ab2) made against a CENP-A peptide (A) or a fusion protein (B) (see Fig. S1). C and D. CENP-C (C) and NDC80 (D) are also reduced on Ascaris chromosome regions that remain at the metaphase plate and will be eliminated. E. IHC staining of microtubules and CENP-A in a 2-cell Ascaris embryo at metaphase illustrates that CENP-A extends along the length and is localized asymmetrically to the centrosome side of the chromosomes. F. IHC staining of CENP-A in a 1-cell Parascaris univalens embryo during prometaphase. Note that CENP-A is reduced on the long arms (arrows) of P. univalens chromosomes just before DNA elimination. These regions will be lost in subsequent cell divisions during DNA elimination, whereas the central chromosome region with CENP-A will form many new retained chromosomes (see G). G. IHC staining of CENP-A in a 4-cell embryo in P. univalens undergoing DNA elimination. Note CENP-A only stains the euchromatin regions (yellow arrows) and the retained genomic regions that form many new chromosomes (white arrows). The size of Ascaris and P. univalens embryos are ~ 70 × 45 μm and ~ 50 × 50 μm, respectively.