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Fig. 2. ARF6 is not required for central spindle or contractile ring formation. (A-E) Time-lapse images of Pav-GFP (A-C) and Sqh-GFP (D,E) during cytokinesis in control (A,D), arf6 (B,E) and chic^13E mutant spermatocytes (C). Times are minutes:seconds after anaphase onset (min AA). Scale bars: 5 µm. (A) Control, Pav-GFP accumulates at the central spindle during anaphase B (arrowhead, 03:29). Central spindle microtubules labelled with Pav-GFP bundle and compact into a dense midbody (arrowheads, 09:14-59:54). (B) arf6¹, anaphase B Pav-GFP central spindle accumulation occurs (arrowhead, 06:54). Pav-GFP-labelled microtubules bundle (arrowhead, 15:20), a cleavage furrow initiates, but central spindle Pav-GFP signal declines (arrowhead, 24:21). After furrow regression, only a tiny amount of Pav-GFP remains at the cortex (arrowhead 50:21). (C) chic^13E, anaphase B Pav-GFP central spindle accumulation occurs (arrowhead, 03:00, 06:54), but no furrow is initiated. 17:30 min AA: very little Pav-GFP remains at the central spindle. (D) Control, Sqh-GFP transfers to the cortex (arrowhead, 04:55), accumulates at the future cleavage furrow site (arrowheads, 07:05, 08:11) which then invaginates (arrowhead, 22:14). (E) arf6³, Sqh-GFP transfers to the cortex (arrowhead, 04:58) concentrating at the future cleavage furrow site (arrowheads 07:01, 14:09). Sqh-GFP remains at the cortex during and after regression (arrowhead, 25:22). Genotypes: w; arf6¹/CyO; Pav-GFP/TM6B (A), w; arf6¹/arf6¹; Pav-GFP/TM6B (B), chic^13E/chic^13E; Pav-GFP/TM6B (C), y w sqh^AX3; +; P (w+ sqh-gfp) (D), y w sqh^AX3; arf6³/arf6³; P (w+ sqh-gfp) (E).

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