Cortical F-actin and cell shape defects in spectrin mutant PECs are rescued by inactivation of cpa. The z-projections at apical domain (A and B) and AJs (C–E), respectively. (A and A’) A Triton X-100–permeabilized pupal eye disc containing GFP-positive MARCM clones with cpa RNAi was stained for phalloidin. Note the increased cortical F-actin level in cpa mutant PECs. Quantification of phalloidin membrane intensity is shown in F. (B and B’) Similar to A and A’ except that MARCM clones with RNAi of both cpa and α-spec were examined. Note the modestly increased cortical F-actin level in the mutant PECs. Quantification of phalloidin membrane intensity is shown in F. (C–C”’) A pupal eye disc containing GFP-positive MARCM clones with cpa RNAi was stained for p-MLC (white, C’) and DE-cad (red, C”). Note the normal p-MLC level in the mutant PECs. Also note the normal apical area of cpa mutant 1° PEC (arrowhead, C”) compared with the wild-type 1° PEC (arrow, C”) in the same mosaic ommatidium. Quantification of apical area of 1° PEC is shown in G. (D–D”’) Similar to C–C’” except that MARCM clones with RNAi of both cpa and α-spec were examined. Note the increased p-MLC level in the mutant PECs. Also note the dramatic decrease in apical area in the mutant 1° PEC (arrowhead, D”) compared with the wild-type 1° PEC (arrow, D”) in the same mosaic ommatidium. Quantification of apical area of 1° PEC is shown in G. (E–E”’) Similar to D–D’” except that the eye disc was treated by the Rok inhibitor Y27632 for 1 h before fixation. Note the similar p-MLC level inside and outside the mutant clones (E’). Also note the increased apical area in the mutant 1° PEC (arrowhead, E”) compared with the wild-type 1° PEC (arrow, E”) in the same mosaic ommatidium. Quantification of apical area of 1° PEC is shown in G. (F) Normalized mean phalloidin membrane intensity analyzed in A–B. Data are means ± SEM (n ≥ 20 cells, representative of five animals). ****, P < 0.0001; **, P < 0.01 (Student’s t test, all compared with wild type). (G) Quantification of 1° PECs apical area of the indicated genotypes analyzed in C–E (means ± SEM, n = 15 for each genotype). ****, P < 0.0001. Scale bars, 5 µm.
Figure 7.

Cortical F-actin and cell shape defects in spectrin mutant PECs are rescued by inactivation of cpa. The z-projections at apical domain (A and B) and AJs (C–E), respectively. (A and A’) A Triton X-100–permeabilized pupal eye disc containing GFP-positive MARCM clones with cpa RNAi was stained for phalloidin. Note the increased cortical F-actin level in cpa mutant PECs. Quantification of phalloidin membrane intensity is shown in F. (B and B’) Similar to A and A’ except that MARCM clones with RNAi of both cpa and α-spec were examined. Note the modestly increased cortical F-actin level in the mutant PECs. Quantification of phalloidin membrane intensity is shown in F. (C–C”’) A pupal eye disc containing GFP-positive MARCM clones with cpa RNAi was stained for p-MLC (white, C’) and DE-cad (red, C”). Note the normal p-MLC level in the mutant PECs. Also note the normal apical area of cpa mutant 1° PEC (arrowhead, C”) compared with the wild-type 1° PEC (arrow, C”) in the same mosaic ommatidium. Quantification of apical area of 1° PEC is shown in G. (D–D”’) Similar to C–C’” except that MARCM clones with RNAi of both cpa and α-spec were examined. Note the increased p-MLC level in the mutant PECs. Also note the dramatic decrease in apical area in the mutant 1° PEC (arrowhead, D”) compared with the wild-type 1° PEC (arrow, D”) in the same mosaic ommatidium. Quantification of apical area of 1° PEC is shown in G. (E–E”’) Similar to D–D’” except that the eye disc was treated by the Rok inhibitor Y27632 for 1 h before fixation. Note the similar p-MLC level inside and outside the mutant clones (E’). Also note the increased apical area in the mutant 1° PEC (arrowhead, E”) compared with the wild-type 1° PEC (arrow, E”) in the same mosaic ommatidium. Quantification of apical area of 1° PEC is shown in G. (F) Normalized mean phalloidin membrane intensity analyzed in A–B. Data are means ± SEM (n ≥ 20 cells, representative of five animals). ****, P < 0.0001; **, P < 0.01 (Student’s t test, all compared with wild type). (G) Quantification of 1° PECs apical area of the indicated genotypes analyzed in C–E (means ± SEM, n = 15 for each genotype). ****, P < 0.0001. Scale bars, 5 µm.

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