Figure 8.
Overexpression of ex or the actin capping protein cpb partially rescues cell migration and polarity defects. (A–J) Confocal micrographs of Crb or F-actin (red) staining in egg chambers harboring GFP-labeled MARCM clones (green) of different genotypes. TO-PRO-3 (blue) labels all nuclei. Scale bars are 25 µm (insets, 10 µm). The stage of egg chamber development is as indicated, with dotted lines showing the position of overlying centripetal follicle cells in stage 9 chambers. BCs are indicated with arrows. (A) Control showing normal distribution of Crb at contacts between the BCs inside the cluster. (B) Control showing normal cortical distribution of F-actin around the outer membrane of the cluster. (C)exe1 clones showing partial disruption of Crb. (D) F-actin polarization is also partially impaired in exe1 BCs, with some F-actin visible at inner junctions of the migrating clusters. (E) Crb is redistributed away from inner junctions to the cortex of the cluster in not1 clones. (F) F-actin is found distributed on inner junctions of not1 clusters between BCs. (G) The disruption of Crb localization in not1 clones is partially rescued by overexpression of ex (not1 ex+). (H) F-actin also is more normally polarized in not1 ex+ BCs, although some weak staining is also evident between BC–BC junctions. (I) Overexpression of cpb weakly restores some Crb distribution in not1 cells (not1 cpb+). (J) F-actin is displaced from BC junctions inside the not1 cpb+ clusters. (K) Quantification of mean percentage of Crb staining at the back, middle, and front (BMF) of the cluster (area under curve measurements) derived from n > 5 egg chambers. Results of two-way ANOVA comparisons of mean ratio of Crb staining in the middle of the cluster are shown above. (L) Quantification of mean percentage of F-actin staining at the BMF of the cluster (area under curve measurements) derived from n > 6 egg chambers. Two-way ANOVA comparisons of mean ratio of F-actin staining in the middle of the cluster are also shown. (M) Plots showing overall mean migration ± SEM derived from means of each replicate (n ≥ 15 egg chambers) of the indicated genotypes. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by unpaired two-tailed Student’s t test. st., stage.

Overexpression of e x or the actin capping protein cp b partially rescues cell migration and polarity defects. (A–J) Confocal micrographs of Crb or F-actin (red) staining in egg chambers harboring GFP-labeled MARCM clones (green) of different genotypes. TO-PRO-3 (blue) labels all nuclei. Scale bars are 25 µm (insets, 10 µm). The stage of egg chamber development is as indicated, with dotted lines showing the position of overlying centripetal follicle cells in stage 9 chambers. BCs are indicated with arrows. (A) Control showing normal distribution of Crb at contacts between the BCs inside the cluster. (B) Control showing normal cortical distribution of F-actin around the outer membrane of the cluster. (C)exe1 clones showing partial disruption of Crb. (D) F-actin polarization is also partially impaired in exe1 BCs, with some F-actin visible at inner junctions of the migrating clusters. (E) Crb is redistributed away from inner junctions to the cortex of the cluster in not1 clones. (F) F-actin is found distributed on inner junctions of not1 clusters between BCs. (G) The disruption of Crb localization in not1 clones is partially rescued by overexpression of ex (not1 ex+). (H) F-actin also is more normally polarized in not1 ex+ BCs, although some weak staining is also evident between BC–BC junctions. (I) Overexpression of cpb weakly restores some Crb distribution in not1 cells (not1 cpb+). (J) F-actin is displaced from BC junctions inside the not1 cpb+ clusters. (K) Quantification of mean percentage of Crb staining at the back, middle, and front (BMF) of the cluster (area under curve measurements) derived from n > 5 egg chambers. Results of two-way ANOVA comparisons of mean ratio of Crb staining in the middle of the cluster are shown above. (L) Quantification of mean percentage of F-actin staining at the BMF of the cluster (area under curve measurements) derived from n > 6 egg chambers. Two-way ANOVA comparisons of mean ratio of F-actin staining in the middle of the cluster are also shown. (M) Plots showing overall mean migration ± SEM derived from means of each replicate (n ≥ 15 egg chambers) of the indicated genotypes. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by unpaired two-tailed Student’s t test. st., stage.

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