Figure S3.
Lam-depleted border cells specify and express key border cell genes. (A–C) Representative images of stage 9 border cells stained with the indicated markers in control (A) and Lam1RNAi (B) and Lam2RNAi (C). Left: merged images. Right: grayscale images of F-actin, E-cadherin, and singed staining. (D) Plot of individual (dots) and average ± SEM (bars) cluster circularity, 1 d at 29°C. One-way ANOVA with Tukey post-hoc was performed. n = 27 (w), 63 (Lam1), and 49 (Lam2) clusters. (E) Plot of average ± SEM numbers of border cells in a cluster, 1 d at 29°C. A Krusal–Wallis test was performed. (F and G) Mean F-actin (F) and E-cadherin (G) levels for each cluster (dots) and the average ± SEM (bars). KD condition: 1 d at 29°C (see also Fig. S5 for 3-d F-actin analysis). n = 27 (w), 63 (Lam1), and 49 (Lam2) clusters. One-way ANOVA with Tukey post-hoc was performed for each plot. (H) Clonal analysis of the ratio of mean peripheral E-cadherin in a LamRNAi clone divided by the mean of a wildtype clone. KD condition: 3 d at 29°C. The middle bars show the mean ± SEM. A Wilcoxon test was performed. n = 61 (Lam1) and 36 (Lam2) clusters. (I and J) Confocal images of STAT activity reporter (10XSTATGFP) surrounding polar cells (asterisks) for the indicated conditions. LamRNAi line shown: Lam1RNAi. KD condition: 1 d at 29°C. (K) Plot showing individual (dots) and mean ± SEM (bars) measures of STAT fluorescence normalized to the mean of the control. n = 27 (w), 45 (Lam1), and 13 (Lam2) clusters. A Kruskal–Wallis test was used for statistical testing. (L) A border cell cluster expressing hsFlpAyGal4 UAS-GFP UAS-LamRNAi clones (GFP+ cells marked with arrowheads) and wildtype clones with the indicated markers a merge of channels. (L′) Grayscale image of notch responsive element RFP. (M) Plot showing the fluorescence intensity (F.I.) of the nuclear notch responsive element relative to wildtype clones in the cluster. KD condition: 3 d at 29°C. Middle bars show the mean ± SEM. n = 43 clusters. A Wilcoxon test was used to test for statistical upregulation, P = 0.06, ns. Scale bars: 10 µm. Genotypes and experimental replicates reported in Table S2.

Lam-depleted border cells specify and express key border cell genes. (A–C) Representative images of stage 9 border cells stained with the indicated markers in control (A) and Lam1RNAi (B) and Lam2RNAi (C). Left: merged images. Right: grayscale images of F-actin, E-cadherin, and singed staining. (D) Plot of individual (dots) and average ± SEM (bars) cluster circularity, 1 d at 29°C. One-way ANOVA with Tukey post-hoc was performed. n = 27 (w), 63 (Lam1), and 49 (Lam2) clusters. (E) Plot of average ± SEM numbers of border cells in a cluster, 1 d at 29°C. A Krusal–Wallis test was performed. (F and G) Mean F-actin (F) and E-cadherin (G) levels for each cluster (dots) and the average ± SEM (bars). KD condition: 1 d at 29°C (see also Fig. S5 for 3-d F-actin analysis). n = 27 (w), 63 (Lam1), and 49 (Lam2) clusters. One-way ANOVA with Tukey post-hoc was performed for each plot. (H) Clonal analysis of the ratio of mean peripheral E-cadherin in a LamRNAi clone divided by the mean of a wildtype clone. KD condition: 3 d at 29°C. The middle bars show the mean ± SEM. A Wilcoxon test was performed. n = 61 (Lam1) and 36 (Lam2) clusters. (I and J) Confocal images of STAT activity reporter (10XSTATGFP) surrounding polar cells (asterisks) for the indicated conditions. LamRNAi line shown: Lam1RNAi. KD condition: 1 d at 29°C. (K) Plot showing individual (dots) and mean ± SEM (bars) measures of STAT fluorescence normalized to the mean of the control. n = 27 (w), 45 (Lam1), and 13 (Lam2) clusters. A Kruskal–Wallis test was used for statistical testing. (L) A border cell cluster expressing hsFlpAyGal4 UAS-GFP UAS-LamRNAi clones (GFP+ cells marked with arrowheads) and wildtype clones with the indicated markers a merge of channels. (L′) Grayscale image of notch responsive element RFP. (M) Plot showing the fluorescence intensity (F.I.) of the nuclear notch responsive element relative to wildtype clones in the cluster. KD condition: 3 d at 29°C. Middle bars show the mean ± SEM. n = 43 clusters. A Wilcoxon test was used to test for statistical upregulation, P = 0.06, ns. Scale bars: 10 µm. Genotypes and experimental replicates reported in Table S2.

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