Figure 2.
B-type lamin promotes border cell delamination. (A) Schematic of the nuclear envelope and the two Drosophila lamin filament networks made of Lam and LamC proteins. (B and C) Images of fixed border cell clusters stained with Lam (B) and LamC (C). Scale bar: 10 μm. (D–G) Images of fixed stage 10 egg chambers for the indicated RNAi and antibody stains. Flies were incubated for 1 d at 29°C. The oocyte is always shown on the right and the white box marks the border cell cluster. (H) Schematic of migration index used to score stage 10 egg chambers. (I and J) Plots showing migration indexes for indicated conditions. n = number of egg chambers. Migration index: magenta: 0%, green: <50%, blue: >50%, and gray: 100%. (I) Statistical test, 1 d at 29°C: Fisher’s exact test with Bonferroni correction yields significant difference in percentage with complete migration, wRNAi versus Lam1RNAi, P < 0.0002, and wRNAi versus Lam2RNAi, P < 0.0002. Statistical test, 3 d at 29°C: Fisher’s exact test with Bonferroni correction for difference in percentage with complete migration, wRNAi versus Lam1RNAi, P < 0.0002, and wRNAi versus Lam2RNAi, P < 0.0002. (J) Statistical test 1 d at 29°C: Fisher’s exact test for difference in percentage with complete migration in wRNAi versus LamCRNAi 1 d, P = 0.08, and Fisher’s exact test wRNAi versus LamCRNAi 3 d yields P = 0.3. (K and L) Maximum projections of time frames from time-lapse series of stage 9 egg chambers expressing slbo4xPHEGFP to mark border cell membranes in control (wRNAi, K) and Lam-depleted border cells (Lam1RNAi, L) after 1 d at 29°C. Time points relative to the start of imaging: magenta = 0 min, yellow = 33 min, green = 54 min, blue = 108 min. Arrowheads mark protrusions. (M) Cluster speed of individual (dots) and mean ± SEM (bars) in control and Lam-depleted clusters. Black outlines: border cell clusters that did not move away from the anterior during the imaging session. Kruskal–Wallis Test. (N) Individual (dots) and mean ± SEM directionality index of clusters measured as total path traveled/net distance. Kruskal–Wallis Test. (M and N)n = 16 (w), 8 (Lam1), and 11 (Lam2) egg chambers. (O) Individual and mean ± SEM speed of delaminated clusters after 3 d incubation at 29°C; n = 6 (w), 5 (Lam1), and 6 (Lam2) egg chambers. One-way ANOVA followed by Tukey post-hoc was performed for statistical testing. Genotypes and experimental replicates are reported in Table S2.

B-type lamin promotes border cell delamination. (A) Schematic of the nuclear envelope and the two Drosophila lamin filament networks made of Lam and LamC proteins. (B and C) Images of fixed border cell clusters stained with Lam (B) and LamC (C). Scale bar: 10 μm. (D–G) Images of fixed stage 10 egg chambers for the indicated RNAi and antibody stains. Flies were incubated for 1 d at 29°C. The oocyte is always shown on the right and the white box marks the border cell cluster. (H) Schematic of migration index used to score stage 10 egg chambers. (I and J) Plots showing migration indexes for indicated conditions. n = number of egg chambers. Migration index: magenta: 0%, green: <50%, blue: >50%, and gray: 100%. (I) Statistical test, 1 d at 29°C: Fisher’s exact test with Bonferroni correction yields significant difference in percentage with complete migration, wRNAi versus Lam1RNAi, P < 0.0002, and wRNAi versus Lam2RNAi, P < 0.0002. Statistical test, 3 d at 29°C: Fisher’s exact test with Bonferroni correction for difference in percentage with complete migration, wRNAi versus Lam1RNAi, P < 0.0002, and wRNAi versus Lam2RNAi, P < 0.0002. (J) Statistical test 1 d at 29°C: Fisher’s exact test for difference in percentage with complete migration in wRNAi versus LamCRNAi 1 d, P = 0.08, and Fisher’s exact test wRNAi versus LamCRNAi 3 d yields P = 0.3. (K and L) Maximum projections of time frames from time-lapse series of stage 9 egg chambers expressing slbo4xPHEGFP to mark border cell membranes in control (wRNAi, K) and Lam-depleted border cells (Lam1RNAi, L) after 1 d at 29°C. Time points relative to the start of imaging: magenta = 0 min, yellow = 33 min, green = 54 min, blue = 108 min. Arrowheads mark protrusions. (M) Cluster speed of individual (dots) and mean ± SEM (bars) in control and Lam-depleted clusters. Black outlines: border cell clusters that did not move away from the anterior during the imaging session. Kruskal–Wallis Test. (N) Individual (dots) and mean ± SEM directionality index of clusters measured as total path traveled/net distance. Kruskal–Wallis Test. (M and N)n = 16 (w), 8 (Lam1), and 11 (Lam2) egg chambers. (O) Individual and mean ± SEM speed of delaminated clusters after 3 d incubation at 29°C; n = 6 (w), 5 (Lam1), and 6 (Lam2) egg chambers. One-way ANOVA followed by Tukey post-hoc was performed for statistical testing. Genotypes and experimental replicates are reported in Table S2.

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