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Fig. 3. LIS-1 distribution in early embryos. (A-I) Wild-type embryos stained with antibodies against LIS-1 and {alpha}-tubulin; upper panels show LIS-1 staining, lower panels the merge of LIS-1 (red), {alpha}-tubulin (green), and Hoechst counterstain to view DNA (blue). Bar, 10 µm. (A-D) One-cell stage. (A) Early prophase; LIS-1 is excluded from pronuclei. (B) Pronuclear meeting; LIS-1 is no longer excluded from pronuclei. (C) Late prometaphase; arrow points to enriched LIS-1 in the vicinity of chromosomes, arrowheads to LIS-1 depletion at the aster centres. (D) Telophase; arrow indicates LIS-1 enrichment at the ingressing central cortex, arrowheads regions of LIS-1 enrichment in the vicinity of microtubule asters. Square 1 indicates an area of high microtubule density, square 2 an area of low microtubule density. Note that LIS-1 staining is more intense in square 1. (E) Two-cell stage; note enrichment of LIS-1 on the P1 spindle (arrowhead) and the cell cortex between AB and P1 (arrow). (F) Four-cell stage; arrowheads point to LIS-1 enrichment in ABa and ABp prophase nuclei. (G,H) Magnified views of the nucleus in the AB blastomere of a two-cell-stage embryo (G) and of the region containing chromosomes in a prometaphase one-cell-stage embryo (H). Arrows point to LIS-1 enrichment at the nuclear periphery (G) and in the vicinity of chromosomes (H). Bar, 2 µm. (I) Three-fold magnification of P1 spindle in panel E. Shown are immunostaining of LIS-1, {alpha}-tubulin and the merge of LIS-1 (red), {alpha}-tubulin (green) and DNA (blue). Arrow points to an apparent microtubule plus end that does not colocalize with a focus of LIS-1, arrowhead to one that does (see also corresponding Movie 8; supplementary material, http://jcs.biologists.org/cgi/content/full/117/19/4571/DC1). (J,K) GFP–LIS-1 transgenic two-cell-stage (J) and four-cell-stage (K) embryos stained with antibodies against GFP and {alpha}-tubulin; upper panels show GFP staining, lower panels the merge of GFP (red), {alpha}-tubulin (green), and Hoechst counterstain to view DNA (blue). Images are projections of up to three consecutive 1 µm confocal slices.





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