XB-IMG-77857
Xenbase Image ID: 77857
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Fig. 3. otx2-GR can activate retinoic acid-insensitive XCG expression in whole embryos.
(A) Experimental scheme. Albino embryos were injected with otx2-GR RNA at the two-cell stage. Injected embryos were incubated alone or in dexamethasone (dex) and/or retinoic acid (RA) for 5 hours beginning at blastula (stage 9) or mid-gastrula (stage 11). Arrows indicate the stage treatment was initiated, triangles indicate the stage of harvest.
(B) XCG induction in whole embryos. Embryos injected in one blastomere with 50 pg otx2-GR RNA were left untreated or incubated in dex and/or RA from stage 9 to 11 (a, b) or stage 11 to 15 (c-f). XCG expression was analyzed by in situ hybridization. Open arrowheads indicate endogenous XCG, solid arrowheads mark ectopic XCG. Anteroventral view. (a) untreated; (b) +dex st. 9; (c) untreated; (d) +dex st. 11; (e) +RA st. 11; (f) +RA+dex st. 11.
(C) Western blot analysis of whole embryos. Embryos injected with 50 pg MT-otx2-GR RNA in both blastomeres and prepared as in B were lysed in pools of five for western blot analysis with an anti-myc antibody. One embryo equivalent of protein was analyzed per lane. Ponceau S staining indicated that all lanes were loaded equally (data not shown). Lane 1, untreated; lane 2, +RA st. 9; lane 3, +dex st. 9; lane 4, +RA+dex st. 9; lane 5, untreated; lane 6, +RA st. 11; lane 7, +dex st. 11; lane 8, +RA+dex st. 11. Image published in: Gammill LS and Sive H (1997) Copyright © 1997. 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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