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. 2019 May 22;15(5):e1008083. doi: 10.1371/journal.pgen.1008083

Fig 2. Loss of dpix or git leads to ectopic cell division in multilayered tissues, phenocopies loss of pak, and disrupts egg chamber shape, and cell shape.

Fig 2

(A-C’) Maximum intensity projections of wild-type (wt) (A-A’), git (B-B’) and dpix (C-C’) stage 8 egg chambers visualised with DAPI (blue) and stained with phospho-Histone H3 (PHH3) (orange). (A, B, C) show whole egg chambers, and (A’, B’, C’) show enlargements of the posterior region. Scale bars are 50 μm. (D) Quantification of the frequency of PHH3 positive cells in stage 7–9 egg chambers for the indicated genotypes. (E-I) Wild-type egg chamber and mutant egg chambers displaying side-by-side egg chamber fusion defects. Genotypes are: wt (E); git (F); dpix (G); dpix, git (H); pak (I). Scale bars 20 μm. (J) Schematic of the dPix-Git complex highlighting the conserved interaction with Pak (via the SH3 domain of dPix), and tension dependent basal localisation at focal adhesions (via Git’s Paxillin binding domain). (K) Scatter plot of egg chamber width against length for wt (green), git (orange) and dpix (blue) genotypes between stages 3 and 10. (L) Box plot representation and statistical comparison of egg chamber aspect ratio between stages 4–8 of egg chamber development. Genotypes: w = wild-type; g = git; p = dpix. Statistical tests are ANOVA with post hoc Tukey’s test, and significance is: * = p < 0.05; *** = p < 0.001; **** = p < 0.0001; ns = not significant. (M-O) Examples of stage 8 egg chambers from wt (M), git (N), and dpix (O) egg chambers, with aspect ratio (AR) displayed. Visualised with DAPI (blue). Scale bars 50 μm. (P-Q) Cortical actin stains at the basal surface of wild-type and git mutant stage 7–8 epithelia. Scale bars 10 μm. (R) Quantification of mean cell eccentricity in wild-type, dpix and git mutant stage 7–8 egg chambers. Error bars are standard deviation. Statistical test is ANOVA with post hoc Dunnett’s test. Each ‘n’ represents a separate egg chamber, with a continuous area of at least 40 cells measured per egg chamber. Sample sizes are: wt, n = 8; dpix, n = 5; git, n = 4. Significance: * = p < 0.05; ** = p < 0.01. (S, T) Choropleth map of wild-type and git epithelia, with each cell coloured according to the degree of eccentricity.