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. Author manuscript; available in PMC: 2017 May 10.
Published in final edited form as: Cell Rep. 2016 Apr 28;15(6):1125–1133. doi: 10.1016/j.celrep.2016.04.014

Figure 4. Fat2 regulates rotation initiation and MT chirality.

Figure 4

(A) ImSAnE unrolling of basal surface of fat2-depleted ovariole, showing regions 2b through st. 3 stained with phalloidin and anti-acTub. (B) Nematic order shows MT and actin alignment resemble WT until st. 2, but become disrupted following st. 3. (C) Rotation speeds at st. 7-8 of follicles depleted of fat2 by RNAi at specified stages. Follicles depleted of fat2 either prior to or following st. 3 fail to rotate. (D) Aspect ratios of eggs from conditional depletion of fat2, showing strong early and late requirements for egg shape. (E) Egg shapes from fat2-RNAi-mild genetic interaction tests. Heterozygosity for either fat2 or CLASP enhances the round egg phenotype. (F) Still frame from live imaging of MT +end growth in WT st. 1 follicle. (G) Quantification of EB1 growth bias in WT and fat2-depleted st. 1 follicle (0=unbiased direction of growth, 1=fully concordant direction of growth), shows that significant MT growth bias in WT is lost when fat2 is depleted. (H) Population-level MT growth biases at st. 1 and follicle rotation directions at st. 6 show similar proportions of chiralities. (I) Live imaging of EB1-GFP expressing follicles during st.1 to st. 2 transition reveals rotation with a chirality opposite to that of MT growth bias.