Analysis of ommatidia size, cell polarity, and Ena expression/function in pupal retina.(A–B”’) A pupal eye disc containing GFP-positive MARCM clones with α-spec RNAi was stained for PAR-3 (green) and Dlg (red), which mark the apical and lateral domain of epithelia cells, respectively. (B–B”’) A vertical section shown through the eye disc in A–A”’ in which the position of the vertical section is indicated by a straight dotted line in A. A’ and A” show z-projections around cell apical and lateral plane, respectively. Note the normal distribution of PAR-3 and Dlg in α-spec mutant PECs (arrowheads) compared with wild-type PECs (arrows). Also note α-spec mutant PECs exhibit increased lateral areas than wild-type cells (compare arrowheads and arrows in A” and B”). Quantification of ommatidium size at lateral position is shown in E. (C–D’) A pupal eye disc containing GFP-positive MARCM clones with α-spec RNAi was stained for DE-cad. D and D’ are magnified view of the dashed-line circled wild-type and mutant ommatidia in C’. Note that α-spec mutant ommatidium (green label in C’ and D’) exhibit increased apical size compared with wild-type ommatidium (white label in C’ and D). Quantification of ommatidium size at apical position is shown in E. (E) Normalized mean ommatidium size at apical and lateral position for the indicated genotypes. Data are means ± SEM (n ≥ 15 ommatidia, representative of five animals). ****, P < 0.0001. (F–G’) Pupal eye discs containing GFP-positive β-specC mutant MARCM clones were stained for DE-cad (red, in F and F’), or Dlg (red, in G and G’). β-specC mutant PECs (arrowhead in F) have similar apical size as wild-type PECs (arrow in F) but exhibit increased lateral size (arrowheads in G) compared with wild-type PECs (arrows in G). Quantification of ommatidium apical and lateral size is shown in H. (H) Normalized mean ommatidium apical and lateral size for the indicated genotypes. Data are means ± SEM (n ≥ 15 ommatidia, representative of five animals). ****, P < 0.0001. N.S., no significance. (I and I’) A pupal eye disc containing GFP-positive MARCM clones with α-spec RNAi was stained for Ena (red), showing similar Ena protein level and localization inside and outside the mutant clones. Quantification of Ena membrane intensity is shown in J. (J) Mean Ena membrane intensity analyzed in I. Data are means ± SEM (n ≥ 20 cells, representative of five animals). N.S., no significance. (K and K’) A Triton X-100–permeabilized pupal eye disc containing GFP-positive ena46 mutant MARCM clones was stained for phalloidin (red). Note the similar cortical F-actin level inside and outside the mutant clones. Quantification of phalloidin membrane intensity is shown in L. (L) Mean phalloidin membrane intensity analyzed in K. Data are means ± SEM (n ≥ 20 cells, representative of five animals). Scale bars, 5 µm. N.S., no significance.
Figure S2.

Analysis of ommatidia size, cell polarity, and Ena expression/function in pupal retina. (A–B”’) A pupal eye disc containing GFP-positive MARCM clones with α-spec RNAi was stained for PAR-3 (green) and Dlg (red), which mark the apical and lateral domain of epithelia cells, respectively. (B–B”’) A vertical section shown through the eye disc in A–A”’ in which the position of the vertical section is indicated by a straight dotted line in A. A’ and A” show z-projections around cell apical and lateral plane, respectively. Note the normal distribution of PAR-3 and Dlg in α-spec mutant PECs (arrowheads) compared with wild-type PECs (arrows). Also note α-spec mutant PECs exhibit increased lateral areas than wild-type cells (compare arrowheads and arrows in A” and B”). Quantification of ommatidium size at lateral position is shown in E. (C–D’) A pupal eye disc containing GFP-positive MARCM clones with α-spec RNAi was stained for DE-cad. D and D’ are magnified view of the dashed-line circled wild-type and mutant ommatidia in C’. Note that α-spec mutant ommatidium (green label in C’ and D’) exhibit increased apical size compared with wild-type ommatidium (white label in C’ and D). Quantification of ommatidium size at apical position is shown in E. (E) Normalized mean ommatidium size at apical and lateral position for the indicated genotypes. Data are means ± SEM (n ≥ 15 ommatidia, representative of five animals). ****, P < 0.0001. (F–G’) Pupal eye discs containing GFP-positive β-specC mutant MARCM clones were stained for DE-cad (red, in F and F’), or Dlg (red, in G and G’). β-specC mutant PECs (arrowhead in F) have similar apical size as wild-type PECs (arrow in F) but exhibit increased lateral size (arrowheads in G) compared with wild-type PECs (arrows in G). Quantification of ommatidium apical and lateral size is shown in H. (H) Normalized mean ommatidium apical and lateral size for the indicated genotypes. Data are means ± SEM (n ≥ 15 ommatidia, representative of five animals). ****, P < 0.0001. N.S., no significance. (I and I’) A pupal eye disc containing GFP-positive MARCM clones with α-spec RNAi was stained for Ena (red), showing similar Ena protein level and localization inside and outside the mutant clones. Quantification of Ena membrane intensity is shown in J. (J) Mean Ena membrane intensity analyzed in I. Data are means ± SEM (n ≥ 20 cells, representative of five animals). N.S., no significance. (K and K’) A Triton X-100–permeabilized pupal eye disc containing GFP-positive ena46 mutant MARCM clones was stained for phalloidin (red). Note the similar cortical F-actin level inside and outside the mutant clones. Quantification of phalloidin membrane intensity is shown in L. (L) Mean phalloidin membrane intensity analyzed in K. Data are means ± SEM (n ≥ 20 cells, representative of five animals). Scale bars, 5 µm. N.S., no significance.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal