Figure S2.
Stronger depletion of lamins correlates with higher frequency of migration defects. (A–D) Images of Lam (A and B) and LamC (C and D) for the indicated conditions. (E–G) Plots showing the mean ± SEM percentage of border cells with the indicated depletion (full, partial, or none) of Lam or LamC at the nuclear periphery for the indicated conditions. (E) A Kruskal–Wallis Test was performed to compare means of cells with a full knockdown (KD): wRNAi versus Lam1, P = 0.006 and w versus Lam2 P < 0.0001. (F) Kruskal–Wallis Test was performed to compare those with a full KD: w versus Lam1, P < 0.0001 and w versus Lam2, P < 0.0001. (G) A Kruskal–Wallis Test was performed to compare the mean of cells with a full KD; wRNAi versus LamC RNAi, P < 0.0001. (H) Plot showing the individual and average ± SEM percent of cells with an indicated knockdown in Lam-depleted clusters with complete (n = 36) or incomplete (n = 31) migration. A Kruskal–Wallis test was performed. Data shown are from line Lam2 RNAi after 1 d at 29°C. (I) Migration index of stage 9 egg chambers for the indicated conditions to quantify the percentage that moved away from the anterior (gray). n = number of egg chambers, after 1 d at 29°C. A Fisher’s exact test with a Bonferroni correction to compare percentage with delamination yields P = 0.0003 for wRNAi versus Lam1RNAi and P < 0.0002 for wRNAi versus Lam2RNAi. (J and K) Migration index for the indicated conditions; n = number of egg chambers. A Fisher’s exact test with a Bonferroni correction to compare percentage with complete migration. P values for J: wRNAi versus Lam1RNAi, P < 0.0002; wRNAi versus Lam2RNAi, P < 0.0002. P values for K: wRNAi versus Lam1 RNAi, P = 0.4; wRNAi versus Lam2RNAi, P > 0.9. KD condition: 1 d at 29°C. (L and M) Images of fixed border cells from stage 9 egg chambers stained for LBR with LacZ overexpressed (OE; L) or LBR overexpressed (M) with c306Gal4 for 1 d at 29°C. (L′ and M′) Grayscale images of LBR staining. (N and O) Images of Hoechst (DNA) staining of stage 9 egg chambers overexpressing LacZ (N) or LBR (O) kept at 29°C for 1 d. Scale bars: 10 μm. Arrowhead: elongated nucleus. (P) Plots measuring nuclear shape in nuclear aspect ratio from Hoechst channel of fixed border cell. Each dot represents an individual nucleus, and the middle and error bars represent the mean ± SEM. A Mann–Whitney test was used for statistical testing. N = 3 experimental replicates, n = 112 (Lac), and n = 123 (LBR) nuclei. (Q and R) Plots of migration indexes of stage 10 egg chambers in the noted conditions. n = number of egg chambers counted. A Fisher’s exact test was used to compare percentage with complete migration and yields P < 0.0001 for Q and P = 0.5 for R. Genotypes and experimental replicates reported in Table S2.

Stronger depletion of lamins correlates with higher frequency of migration defects. (A–D) Images of Lam (A and B) and LamC (C and D) for the indicated conditions. (E–G) Plots showing the mean ± SEM percentage of border cells with the indicated depletion (full, partial, or none) of Lam or LamC at the nuclear periphery for the indicated conditions. (E) A Kruskal–Wallis Test was performed to compare means of cells with a full knockdown (KD): wRNAi versus Lam1, P = 0.006 and w versus Lam2 P < 0.0001. (F) Kruskal–Wallis Test was performed to compare those with a full KD: w versus Lam1, P < 0.0001 and w versus Lam2, P < 0.0001. (G) A Kruskal–Wallis Test was performed to compare the mean of cells with a full KD; wRNAi versus LamC RNAi, P < 0.0001. (H) Plot showing the individual and average ± SEM percent of cells with an indicated knockdown in Lam-depleted clusters with complete (n = 36) or incomplete (n = 31) migration. A Kruskal–Wallis test was performed. Data shown are from line Lam2 RNAi after 1 d at 29°C. (I) Migration index of stage 9 egg chambers for the indicated conditions to quantify the percentage that moved away from the anterior (gray). n = number of egg chambers, after 1 d at 29°C. A Fisher’s exact test with a Bonferroni correction to compare percentage with delamination yields P = 0.0003 for wRNAi versus Lam1RNAi and P < 0.0002 for wRNAi versus Lam2RNAi. (J and K) Migration index for the indicated conditions; n = number of egg chambers. A Fisher’s exact test with a Bonferroni correction to compare percentage with complete migration. P values for J: wRNAi versus Lam1RNAi, P < 0.0002; wRNAi versus Lam2RNAi, P < 0.0002. P values for K: wRNAi versus Lam1 RNAi, P = 0.4; wRNAi versus Lam2RNAi, P > 0.9. KD condition: 1 d at 29°C. (L and M) Images of fixed border cells from stage 9 egg chambers stained for LBR with LacZ overexpressed (OE; L) or LBR overexpressed (M) with c306Gal4 for 1 d at 29°C. (L′ and M′) Grayscale images of LBR staining. (N and O) Images of Hoechst (DNA) staining of stage 9 egg chambers overexpressing LacZ (N) or LBR (O) kept at 29°C for 1 d. Scale bars: 10 μm. Arrowhead: elongated nucleus. (P) Plots measuring nuclear shape in nuclear aspect ratio from Hoechst channel of fixed border cell. Each dot represents an individual nucleus, and the middle and error bars represent the mean ± SEM. A Mann–Whitney test was used for statistical testing. N = 3 experimental replicates, n = 112 (Lac), and n = 123 (LBR) nuclei. (Q and R) Plots of migration indexes of stage 10 egg chambers in the noted conditions. n = number of egg chambers counted. A Fisher’s exact test was used to compare percentage with complete migration and yields P < 0.0001 for Q and P = 0.5 for R. Genotypes and experimental replicates reported in Table S2.

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