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. Author manuscript; available in PMC: 2021 Dec 27.

Published in final edited form as: Dev Biol. 2006 Apr 27;297(2):323–339. doi: 10.1016/j.ydbio.2006.04.454

Fig. 5. — Inhibition of DRho kinase phenocopies zip mutants. (A) Cuticle; injected with buffer alone, phase contrast. (B, C) Two examples of resulting denticle pattern defects observed after 120 μM injection of the Rok inhibitor. (B) This example highlights shaping defects as some denticles of row 1 and row 4 exhibit ambiguous shaping or reversals hook (arrows). We tallied these defects collectively as shaping defects and also tallied separately reversals in denticle hooking (see histograms, below). This example also shows some ectopic as well as missing denticles. (C) This example highlights irregularities in alignment along rows, as well as missing denticles and reversals of shaping. (D, E) Embryo mock-injected at stage 12 and then aged and processed at stage 15; Phosphotyrosine (green) and phalloidin (red). Robust ABPs are evident, and cells are aligned (arrows). (E) Magnified view of D, showing protrusions positioned at posterior cell edges. The non-homogeneities in phosphotyrosine staining are due to the initial deposition of cuticle which inhibits antibody penetration. (F, G) Two examples of embryos injected with 120 μM Rok inhibitor at stage 12 and then aged and processed at stage 15; phalloidin staining results on stage 15. Note irregularities in size, shaping and placement of protrusions. The inhibitor dramatically affects the accumulation of phosphotyrosine epitopes, so we cannot use this to reveal cell outlines. Outlines are visible from the phalloidin stain, acquired at a higher gain and presented in grayscale. The normal rectilinear alignment of cells within the denticle field is compromised by Rok inhibition. (H) Magnified view of G shows that several protrusions are misplaced to anterior edges or mid-face of cells (arrows). (I, J) Histograms representing quantification of Rok inhibitor experiments. The recipient embryos were either w¹¹¹⁸ (w−) or embryos expressing Sqh E20 E21, and “Mock” were injected with buffer alone, while inhibitor concentrations are as indicted. (I) The number of larval cuticles scored as wild-type for each condition is as follows: for w recipients, buffer-injected, 34 (39 total scored); at 80 μM inhibitor, 2 (28 total scored); 100 μM, 2 (29 total scored); 120 μM, 3 (27 total scored); 240 μM, 0 (40 total scored). For Sqh {E20,E21} recipients, buffer-injected, 43 (44 total scored); at 80 μM, 15 (32 total scored); 100 μM, 22 (42 total scored); 120 μM, 12 (56 total scored); 240 μM, 0 (40 total scored). (J) Fraction of cuticles scored having either aggregate shaping defects or reversals (tallied separately). Scale bar = 10 μm in panels A, D, F, G; 7 μm in B, C; 3 μm in E, H.

Fig. 5. — Inhibition of DRho kinase phenocopies zip mutants. (A) Cuticle; injected with buffer alone, phase contrast. (B, C) Two examples of resulting denticle pattern defects observed after 120 μM injection of the Rok inhibitor. (B) This example highlights shaping defects as some denticles of row 1 and row 4 exhibit ambiguous shaping or reversals hook (arrows). We tallied these defects collectively as shaping defects and also tallied separately reversals in denticle hooking (see histograms, below). This example also shows some ectopic as well as missing denticles. (C) This example highlights irregularities in alignment along rows, as well as missing denticles and reversals of shaping. (D, E) Embryo mock-injected at stage 12 and then aged and processed at stage 15; Phosphotyrosine (green) and phalloidin (red). Robust ABPs are evident, and cells are aligned (arrows). (E) Magnified view of D, showing protrusions positioned at posterior cell edges. The non-homogeneities in phosphotyrosine staining are due to the initial deposition of cuticle which inhibits antibody penetration. (F, G) Two examples of embryos injected with 120 μM Rok inhibitor at stage 12 and then aged and processed at stage 15; phalloidin staining results on stage 15. Note irregularities in size, shaping and placement of protrusions. The inhibitor dramatically affects the accumulation of phosphotyrosine epitopes, so we cannot use this to reveal cell outlines. Outlines are visible from the phalloidin stain, acquired at a higher gain and presented in grayscale. The normal rectilinear alignment of cells within the denticle field is compromised by Rok inhibition. (H) Magnified view of G shows that several protrusions are misplaced to anterior edges or mid-face of cells (arrows). (I, J) Histograms representing quantification of Rok inhibitor experiments. The recipient embryos were either w¹¹¹⁸ (w−) or embryos expressing Sqh E20 E21, and “Mock” were injected with buffer alone, while inhibitor concentrations are as indicted. (I) The number of larval cuticles scored as wild-type for each condition is as follows: for w recipients, buffer-injected, 34 (39 total scored); at 80 μM inhibitor, 2 (28 total scored); 100 μM, 2 (29 total scored); 120 μM, 3 (27 total scored); 240 μM, 0 (40 total scored). For Sqh {E20,E21} recipients, buffer-injected, 43 (44 total scored); at 80 μM, 15 (32 total scored); 100 μM, 22 (42 total scored); 120 μM, 12 (56 total scored); 240 μM, 0 (40 total scored). (J) Fraction of cuticles scored having either aggregate shaping defects or reversals (tallied separately). Scale bar = 10 μm in panels A, D, F, G; 7 μm in B, C; 3 μm in E, H.