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Fig. 7. Relationship between lis-1 and dynein/dynactin. (A,B) Embryo at the two-to-four cell stage transition expressing GFP–LIS-1 and stained with antibodies against DHC-1 and GFP; upper panels shows DHC-1, middle panel shows GFP, lower panel the merge of GFP (red), DHC-1 (green), and Hoechst counterstain to view DNA (blue). Panel A (approximately 6x10 µm) shows high magnification of the P₁ nucleus during late prophase, panel B (approximately 3x5 µm) a cytoplasmic area from the AB cell in the same embryo. Bottom of diamond points to a focus of GFP–LIS-1 that does not coincide with intense DHC-1 staining, arrow to a focus of DHC-1 that does not coincide with intense GFP–LIS-1 staining, arrowhead to a focus of coincident DHC-1 and GFP–LIS-1 staining. (C,D) Wild-type (C) and lis-1(t1550) (D) two-cell-stage-equivalent embryos stained with antibodies against DHC-1 and -tubulin. Upper panels show DHC-1 staining, lower panels the merge of DHC-1(red), -tubulin (green) and Hoechst counterstain to view DNA (blue). Note that DHC-1 enrichment at the nuclear periphery, in the vicinity of microtubule asters (arrowheads) and at the cell cortex (arrows) are all somewhat diminished in lis-1(t1550) mutant embryos compared with wild-type. In rare lis-1(t1550) embryos, enrichment of DHC-1 at specific subcellular locations appeared to be absent (not shown). Bar, 10 µm. (E-G) dhc-1(RNAi) (E), dnc-1(RNAi) (F) or dnc-2(RNAi) (G) embryos at the second cell cycle stained with antibodies against LIS-1 and -tubulin. Upper panels show anti-LIS-1 staining, lower panel the merge of LIS-1(red), -tubulin (green) and Hoechst counterstain to view DNA (blue). Note that LIS-1 is still present at the cell cortex (arrows), albeit to somewhat lower levels than in the wild-type (compare with Fig. 3D). Note also that LIS-1 enrichment along chromosomes is visible in the dnc-2(RNAi) embryo (G, arrowhead). Moreover, note that LIS-1 becomes trapped in nuclear compartments after the first failed cell division in this particular dhc-1(RNAi) embryo (E); this was not observed in all dhc-1(RNAi) embryos examined, and only rarely after inactivation of dynactin components.

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