The cases where the identity of the defective seam nucleus is ambiguous, as in Figure 6B, were excluded inhibitor supplier from the analysis. We observed defects in all of the seam cells, H0 2, V1 V6 and T, suggesting that failure of cell division affects all the cells in the seam cell linage. However, the frequencies of defects are different between the seam cells. For example, H0 seam cell defect was observed only once in 251 animals scored. The H0 cells are the only cells, from the seam cell lineage, that do not undergo postembryonic division, further confirming the previous findings that the seam cell defect observed in mdf 2 homozygotes is mainly due to postembryonic defects. Similarly, H2, V5 and T cell defects were rarely observed.

In contrast, frequent defects were observed in the six seam cells, H1, V1 V4 and V6 that undergo expansion divi sion to generate an additional six seam cells at L2 and beyond. These data support the findings that seam cell defects likely arise in L2 mdf 2 homozygotes. Further more, we quantified extra seam cell nuclei versus missing seam cell nuclei and, as expected, we observed that reduction of the number of SCM,GFP positive nuclei is a much more common event. Representative images of seam cell reduction due to a failure of cell cycle pro gression of a particular lineage are shown in Figure 6. Together, these data indicate that seam cell defects in the absence of MDF 2 are mainly attributed to cell pro liferation failure at L2 which randomly affects H1, V1 V4 or V6 seam cells.

The seam cell reduction in mdf 2 is not likely to be caused by ced 3 dependent cell death It is possible that the reduction of number of seam cells in mdf 2 worms is caused by cell damage followed by apoptotic cell death. CED 3 is a member of the caspase family of cystein proteases that is required for cell death in C. elegans. To determine whether apoptotic cell death could account for loss of seam cells, we con structed ced 3 unc 26 mdf 2 in which there is no cell death. We found that ced 3 unc 26 mutants do not affect seam cell develop ment, as on average 15. 92 seam cell nuclei were observed in young adults. Further more, we found that ced 3 unc 26 mdf 2 animals had similar numbers of seam cell nuclei as mdf 2, suggesting that ced 3 dependent cell death is unlikely to be responsible for seam cell loss in the tm2190 background.

Absence of FZR 1 enhances sterility of mdf 2 mutants without causing any effect on seam cell development During postembryonic development, seam cell division is regulated at the G1 to S phase progression by a cascade of regulatory factors that include LIN 35 Batimastat Rb, FZR 1 Cdh1, and CKI 1. As LIN 35 and FZR 1 act redundantly to control the G1 to S phase progression, seam cell proliferation appears to be normal in lin 35 and fzr 1 single mutants, while extensive hyperprolifera tion is observed in lin 35, fzr 1 double mutants.