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Fig. 6. LIS-1 targeting to the cell cortex, the nuclear periphery and kinetochores is microtubule-independent. Wild-type (C) or lis-1(1550) embryos rescued by GFP–LIS-1 expression (A,B,D,E) treated with tba-2 (RNAi) and stained with antibodies against LIS-1 and -tubulin; upper panels show LIS-1 staining, lower panels the merge of LIS-1 (red), -tubulin (green), and Hoechst counterstain to view DNA (blue). (A) One-cell-stage embryo prior to pronuclear envelope breakdown; note that GFP–LIS-1 is excluded from pronuclei. (B) One-cell-stage embryo during mitosis; note intense GFP–LIS-1 in the vicinity of condensed chromosomes (arrow). (C) High magnification view of chromosomes in an embryo at a later cell cycle. Embryo posterior is up; bar, 2 µm. Note that LIS-1 enrichment follows the position of chromosomes. (D) Embryo at a later cell cycle; note cortical enrichment of GFP–LIS-1 (arrow). (E) Later-stage embryo during prophase; note enrichment at the nuclear periphery (arrow). Images are projections of up to three consecutive 1 µm confocal slices.

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