Research article

Diverse roles of actin in C. elegans early embryogenesis

Nathalie Velarde, Kristin C Gunsalus and Fabio Piano^*

* Corresponding author: Fabio Piano fp1@nyu.edu

New York University, Department of Biology and Center for Comparative Functional Genomics, 100 Washington Square East, New York, NY 10003, USA

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BMC Developmental Biology 2007, 7:142 doi:10.1186/1471-213X-7-142

Published: 24 December 2007

Additional files

Additional File 1:

Supplementary Figure 1 – ClustalW multiple DNA sequence alignment of conceptually spliced coding sequences for the three embryonic actins with the PCR product (sjj_T04C12.6) used as the template for act-1(RNAi) dsRNA. The top chart shows that all three actins (labeled "act-1", "act-2", and "act-3") show a high percent identity to each other and to the PCR product used for RNAi (labeled "act_1dsRNA"). Very high similarity extends across an extensive portion of all three actin sequences, including several long stretches of 100% identity, indicating that all three embryonic actins are depleted by act-1(RNAi) in this study.

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Additional File 4:

Movie 1 – Embryogenesis in wild type (DIC and GFP::HIS)

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Additional File 5:

Movie 2 – Class I actin(RNAi) (DIC)

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Additional File 6:

Movie 3 – Class II actin(RNAi) (DIC)

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Additional File 7:

Movie 4 – Class III actin(RNAi) (DIC)

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Additional File 8:

Movie 5 – Class IV actin(RNAi) (DIC)

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Additional File 2:

Supplementary Figure 2 – actin(RNAi) affects egg production and fertilization. Young adult hermaphrodites, either untreated [N = 3 for the N2 (WT) strain] or injected with act-1 dsRNA [N = 7 for N2; N = 13 for the PF100 (GFP::MOE) strain], were individually transferred to a new plate every 8 hrs. Plates were then scored for fertilized eggs (characterized by the presence of an eggshell and oval shape) and unfertilized oocytes (characterized by a large endomitotic nucleus and no eggshell). The proportion of unfertilized oocytes and fertilized eggs (Y-axis) was then plotted against the six successive time intervals (X-axis) for each strain and treatment scored (total number of eggs and oocytes scored are given in parentheses). The graph shows that after dsRNA injection, an increasing proportion of oocytes laid over time are unfertilized, and egg production eventually ceases altogether by 48 hrs, whereas WT animals continue to lay fertilized eggs at approximately the same rate.

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Additional File 3:

Supplementary Table 1 – Summary of phenotypic defects in actin(RNAi) embryos. The range of normal and abnormal events observed and their occurrence in the four phenotypic classes of actin-depleted embryos vs. WT embryos is shown. Symbols: (+), observed; (-), not observed; n/a, not applicable.

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Additional File 9:

Movie 6 – Early embryogenesis in GFP::MOE strain

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Movie 7 – par-6(RNAi) of GFP::MOE strain

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Additional File 11:

Movie 8 – par-2(RNAi) of GFP::MOE strain