Morpholinos for splice modificatio

Morpholinos for splice modification

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Supplementary Material

DEV071407 Supplementary Material

Files in this Data Supplement:

  • Supplemental Figure S1 -

    Fig. S1. Quantitation of eye volume, cell count and mitoses. (A) Eye domain absolute volumes (24 hpf) for the three datasets. (B) Absolute volumes of the eye and domains over time. n=3; error bars, s.d. (C) Relative cell count over time (n=3). (D) Cell density over time (n=3). (E,F) Mitosis visualization in 4D datasets. Three-dimensional rendering of nuclei (grayscale) and mitoses (color). (E) Time point t001. Circles, metaphase plates. (F) Time point t002. Circles, daughter nuclei from previous time point; asterisks, new metaphase plates. Dorsal views. Insets show chromatin morphology. Scale bar: 50 ┬Ám.

  • Supplemental Figure S2 -

    Fig. S2. Cells move rapidly during OV elongation. Scatter plot of retina, RPE and lens cell speeds during OV elongation (triangles, 12-14 hpf), as compared with speeds for the following 2 hours (circles, 14-16 hpf). Black bars indicate average speed, which is significantly slower after elongation (P<0.05 for all three tissues). Retina, blue; RPE, green; lens, red.

  • Supplemental Figure S3 -

    Fig. S3. Clustering of tracked cells. Tracked cells were assigned to domains based on final position (24 hpf) and trajectory shape. Volume renderings of tracked nuclei at 12 (A,C,E,G,I,K) and 24 (B,D,F,H,J,L) hpf. (A-F) Retinal nuclear positions. (A,B) Anterior and posterior domains. Dorsal view. (C,D) Central domain. Dorsal view. (E,F) Dorsal and ventral domains. Lateral view. (G-J) RPE nuclear positions. (G,H) Anterior and posterior domains. Dorsal view. (I,J) Central domain. Dorsal view. (K,L) Lens nuclear positions. Anterior and posterior domains.

  • Supplemental Table S1 -
  • Supplemental Table S2 -
  • Movie 1 -

    Movie 1. Time-lapse of eye morphogenesis. Single confocal section from a 4D dataset showing right OV at 12-24 hpf. Dorsal view. Embryo labeled with EGFP-CAAX (membranes, green) and H2A.F/Z-mCherry (nuclei, magenta).

  • Movie 2 -

    Movie 2. Proliferation during OCM. Four-dimensional movie of mitoses and nuclear channel (grayscale). Nuclei were marked at a time point immediately preceding chromosome separation (anaphase) and at anaphase. Mitoses are color-coded blue (dorsal) to red (ventral).

  • Movie 3 -

    Movie 3. Extended evagination (12-17 hpf). Cells are moving from the brain to OV. Trajectories are superimposed on an average projection of the membrane channel (grayscale).

  • Movie 4 -

    Movie 4. OV elongation (12-14 hpf). Pinwheel movement of NR and RPE. Trajectories are superimposed on an average projection of the membrane channel (grayscale).

  • Movie 5 -

    Movie 5. Rim movement (18-24 hpf). Medial layer cells involute around the OV rim, joining the lateral layer. Trajectories are superimposed on an average projection of the membrane channel (grayscale).

  • Movie 6 -

    Movie 6. Position of tracked retinal nuclei at 12 and 24 hpf. Three-dimensional rendering of nuclei and membrane channel (grayscale).

  • Movie 7 -

    Movie 7. Retinal subdomains at 12 and 24 hpf. Domains are segmented based on nuclear position at 24 hpf and trajectory shape. Posterior, yellow; central, blue; anterior, orange; dorsal, green; ventral, red.

  • Movie 8 -

    Movie 8. Retinal (left), RPE (center) and lens (right) trajectories. Three-dimensional rendering of trajectories and membrane channel (grayscale). Retinal trajectories: posterior, yellow; central, blue; anterior, orange; dorsal, green; ventral, red. RPE trajectories: posterior, yellow; central, blue; anterior, red. Lens trajectories: posterior, red; anterior, blue.

  • Movie 9 -

    Movie 9. Central domain merge (14-24 hpf). Posterior (yellow) and anterior (orange) cells move and encircle the central (blue) cells. Trajectories are superimposed on an average projection of the membrane channel.

  • Movie 10 -

    Movie 10. RPE spreading (13.75-24 hpf). Prospective RPE cells spread and surround the NR. Trajectories are superimposed on an average projection of the membrane channel.

  • Movie 11 -

    Movie 11. Position of tracked RPE nuclei at 12 and 24 hpf. Three-dimensional rendering of nuclei and membrane channel (grayscale).

  • Movie 12 -

    Movie 12. RPE subdomains at 12 and 24 hpf. Domains are segmented based on nuclear position at 24 hpf and trajectory shape. Posterior, yellow; central, blue; anterior, red.

  • Movie 13 -

    Movie 13. OV elongation (lens) at 12-14 hpf. Pinwheel movement of lens cells. Trajectories are superimposed on an average projection of the membrane channel.

  • Movie 14 -

    Movie 14. Position of tracked lens nuclei at 12 and 24 hpf. Three-dimensional rendering of nuclei and membrane channel (grayscale).

  • Movie 15 -

    Movie 15. Lens subdomains at 12 and 24 hpf. Domains are segmented based on nuclear position at 24 hpf and trajectory shape. Posterior, red; anterior, blue.