Shaye and Greenwald
Supplementary Information 2
LIN-12(∆DTS)::GFP behaves like LIN-12(+)::GFP in cell fate decisions other than VPC specification.
Alterations in lin-12 activity affect cell fate decisions, including the anchor cell/ventral uterine precursor cell (AC/VU) decision
in the gonad and a sex mesoblast/coelomocyte (SM/cc) decision in the mesoderm of hermaphrodites1. In wild type, the AC/VU
decision produces one AC and one VU, and the SM/cc decision produces two ventral sex mesoblasts and two dorsal
coelomocytes. Loss of lin-12 activity causes 2 ACs (instead of 1 AC) and 2 extra ventral coelomocytes (instead of sex
mesoblasts); elevated or constitutive lin-12 activity results in 0 AC and the absence of dorsal coelomocytes (because they have
been transformed into two extra SMs).
In functional assays, LIN-12(DTS)::GFP behaves like LIN-12(+)::GFP. Both proteins displayed similar ability to complement
the sterility (see Methods) and 2 AC defects associated with the lin-12(n941) null allele. Both proteins also behaved
comparably in assays for cell fate transformations in a lin-12(+) background, suggesting that LIN-12(DTS)::GFP does not
have significant dominant-negative or constitutive activity.
 LIN-12(∆DTS)::GFP rescues the lin-12(0) two anchor cell (2AC) defect.
LIN-12 protein
Genotype
0AC
expressed from arEx array
unc-4(e120); lin-12(n941); arIs51; arEx299
0/19 (0%)
LIN-12(∆DTS)::GFP
unc-4(e120); lin-12(n941); arIs51; arEx300
3/20 (15%)
unc-4(e120); lin-12(n941); arIs51; arEx301
4/24 (17%)
LIN-12(+)::GFP
unc-4(e120); lin-12(n941); arIs51; arEx302
1/35 (3%)
1AC
2AC
15/19 (79%) 4/19 (21%)
13/20 (65%) 4/20 (20%)
18/24 (75%) 2/24 (8%)
31/35 (89%) 3/35 (8%)
 LIN-12(∆DTS)::GFP behaves similarly to LIN-12(+)::GFP in a lin-12(+) background.
AC/VU decision
LIN-12 protein
Genotype
0AC
1AC
2AC
expressed from arEx array
unc-4(e120); arIs51; arEx299
3/20 (15%) 17/20 (85%) 0/20 (0%)
LIN-12(∆DTS)::GFP
unc-4(e120); arIs51; arEx300
4/24 (17%) 20/24 (83%) 0/24 (0%)
unc-4(e120); arIs51; arEx301 LIN-12(+)::GFP
3/20 (15%) 17/20 (85%) 0/20 (0%)
Shaye and Greenwald
Supplementary Information 2
In the absence of transgenes, lin-12(n941) has 2 ACs. The integrated array arIs51 contains pcdh-3::gfp and marks the AC fate
(ref. 2 and X. Karp and I. Greenwald, unpublished observations.)
SM/cc decision
# of posterior coelomocytes
LIN-12 protein
0
2
>2
expressed from arIs77
arIs37; unc-4(e120); arIs77 LIN-12(∆DTS)::GFP
0/37 (0%) 37/37 (100%) 0/37 (0%)
Genotype
To facilitate scoring coelomocytes, we used arIs37, which expresses a secreted form of GFP that is endocytosed by
coelomocytes3.
1.
2.
3.
Greenwald, I. S., Sternberg, P. W. & Horvitz, H. R. The lin-12 locus specifies cell fates in Caenorhabditis elegans. Cell 34, 435-44. (1983).
Pettitt, J., Wood, W. B. & Plasterk, R. H. cdh-3, a gene encoding a member of the cadherin superfamily, functions in epithelial cell
morphogenesis in Caenorhabditis elegans. Development 122, 4149-57. (1996).
Fares, H. & Greenwald, I. Genetic analysis of endocytosis in Caenorhabditis elegans: coelomocyte uptake defective mutants. Genetics 159,
133-45. (2001).