mcf2lb mutants show impaired apical constriction of cells incorporated into rosettes. (A,B) Cellular reconstruction of WT and mcf2lb mutant pLLPs. (C,D) Examples of apically constricted cells from WT (C) and mc2lb mutant (D) pLLPs. The left side panels show the lateral (top-down) view as in panels A and B, and the right panels show the apical/basal views of the cell (cell is virtually turned by 90°). Blue and purple cells marked by the asterisks in panels A and B are also shown in C and D, respectively. (E) Examples of cells that are making contact with multiple points in mcf2lb mutants. Left panel is a lateral (top-down) view; right panel is an apical/basal view. (F) Categorical breakdown of cells in the trailing region of WT (n=258 cells from four pLLPs) and mcf2lb mutants (n=311 cells from four pLLPs). Note that in mcf2lb mutants, there is an increase in the percentage of cells that touch multiple points and an increase in the percentage of cells not incorporated into rosettes. Subsequently, there is a decrease in the percentage of cells incorporated into rosettes. (G) Mean apical constriction index of cells that are incorporated into rosettes in WT (n=169 cells from four pLLPs) and mcf2lb mutants (n=139 cells from four pLLPs). (H) Mean cell volume of cells incorporated into the trailing rosette in WT (n=74 cells from four pLLPs) and mcf2lb mutants (n=91 cells from four pLLPs). (I) Mean apical width of cells incorporated into rosettes in WT (n=169 cells from four pLLPs) and mcf2lb mutants (n=139 cells from four pLLPs). (J) Mean basal width of cells incorporated into rosettes in WT (n=169 cells from four pLLPs) and mcf2lb mutants (n=139 cells from four pLLPs). Note that the apical width is significantly increased in mcf2lb mutants whereas there is no significant difference in the basal width. ***P<0.001 (F: chi-square test; G,I,J: Mann–Whitney U-test; H: unpaired two-tailed t-test). n.s., not significant. Error bars are s.d. Scale bars: 10 μm (A,B and E pLLP); 5 μm (C,D and E individual cells).
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