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Fig. 3. Microtubule organization in sktl mutant oocyte. WT
(A,C,E,G,I,K,N,O) and sktl2.3 mutant (B,D,J,L,M)
Drosophila germline clones and
sktl2.3/sktl
5 mutant
egg chambers (F,H,P,Q). (A-D) Microtubule (MT) network detected with
Khc antibody. In stage 8 sktl2.3 oocytes, MT network
organization is not affected (compare B with A). However, in stage 10
sktl2.3 oocytes, MT organization is disrupted (compare D
with C). MT bundles appear to project from the mislocalized nucleus towards
the center of the cytoplasm (arrowheads in D).
(E,F)Khc-β-gal (red) and DNA (green) staining. In
sktl2.3/sktl
5
oocytes, the posterior pool of Khc-β-gal is lost and it accumulates in
the anterior part of the oocyte (F). The inset in F shows Khc-β-gal
staining alone. (G,H) Dmn-GFP. In
sktl2.3/sktl
5
oocytes, the posterior pool of Dmn-GFP is lost and an accumulation is found in
the most anterior part of the cytoplasm (H). However, cortical- and
nuclear-associated localization are unaffected, even when the nucleus is
mislocalized. (I,J) In both mutant and WT, the centrosome
component DPLP (red) is detected around the oocyte nucleus and as a dot close
to it (arrowhead). The dots surrounding the oocyte correspond to the
centrosomes of the follicular cells. (K-M) Projections of confocal
sections from oocytes immunostained with Spn-F antibody revealing the minus
ends of the MTs. Spn-F is found mislocalized in a characteristic cortical ring
close to the mislocalized nucleus in stage 10 sktl2.3
oocytes (compare M with K), but not in stage 8 (L). (N-Q)MT detection
by anti-Khc. WT (N) and
sktl2.3/sktl
5 (P)
oocytes showing complete depolymerization of MTs after 30 minutes of cold
shock. (O,Q) WT and mutant oocytes after a 10-minute recovery at 25°C. MTs
regrow from the misplaced nucleus (arrowheads in Q). Asterisk, nucleus.