XB-IMG-83475
Xenbase Image ID: 83475
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Figure 4. H3.3 Downregulation Does Not Affect Animal Caps Elongation upon Mesoderm Induction but Leads to Cell Death
(A) Animal Cap assay using MO H3.3 treated embryos. The scheme shows injections in both cells of 2-cell stage embryos with indicated MO [2.5 ng/cell (= 5 ng) or 4.6 ng/cell (= 9.2 ng)] or with Xbra-EnR mRNA [250 pg/cell (= 500 pg)], a dominant-negative form of Xbra as a negative control. After incubation to reach stage 8 in controls, we dissected animal caps in each case for incubation with or without activin (5 ng/ml) for 1.5 hr, and cultured them until sibling embryos reached the neurula stage. For the low MO dose (5 ng), we show an inset of one representative animal cap elongation at the same magnification scale as the other panels. Scale bar: 1 mm.
(B) Analysis of H3.3 morpholino injected embryos throughout development. The scheme shows injections of indicated MO (4.6 ng) in one cell of 2-cell stage embryos. We followed development at 18�C of one control (white box) and five H3.3 morphants with acquisition from the vegetative pole (see also Movie S2). Here are three time points: 64 cells, gastrula, and neurula stage. White arrows indicate the apoptotic cells observed at the end of the gastrulation. Scale bar: 1 mm.
(C) TUNEL assay of MO H3.3 treated embryos. We injected the indicated MO (4.6 ng) in one cell of 2-cell stage embryos and performed a TUNEL assay when controls embryos reached the neurula and tailbud stages. White arrows indicate the TUNEL positive cells in the injected side of H3.3 morphants. The majority of white apoptotic cells comes off the embryos at the beginning of the experimental procedure that involves dechorionization. Scale bar: 1 mm.
See also Figure S5. Image published in: Szenker E et al. (2012) Copyright © 2012. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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