Table 3.
RNA interference of sur-6 PP2A-B, PP2A-C, or PP2A-A
| Genotype
|
dsRNAa or transgenesb
|
Percent Vul or Muv (n)
|
Percent avg. induction (n)
|
|---|---|---|---|
| + | no dsRNA | 0 (many) | 100 (many) |
| + | sur-6 PP2A-B | 19 Vul (21) | 94 (21) |
| + | PP2A-C | 5 Vul (22) | 99 (22) |
| let-60(n1046gf) | no dsRNA | 95 Muv (19) | 173 (19) |
| let-60(n1046gf) | sur-6 PP2A-B | 3 Muv (29) | 101 (29) |
| let-60(n1046gf) | PP2A-C | 70 Muv (23) | 132 (23) |
| let-60(n1046gf) | sur-8 | 15 Muv (33) | 106 (33) |
| let-60(n1046gf) | no dsRNA | 93 Muv (59) | 143 (59) |
| let-60(n1046gf) | sur-6 PP2A-B | 7 Muv (15) | 103 (15) |
| let-60(n1046gf) | PP2A-A | 57 Muv (14) | 125 (14) |
| let-60(n1046gf) | no transgenes | 82 Muv (242) | 175 (23) |
| let-60(n1046gf) | sur-6 PP2A-B | 20 Muv (172) | 110 (48) |
| let-60(n1046gf) | PP2A-C | 65 Muv (82) | N.D. |
In the case of sur-6 PP2A-B, PP2A-C or PP2A-A (but not sur-8), nearly complete embryonic lethality was observed in progeny laid 8–24 hr after double-stranded (ds) RNA injection. Most surviving sur-6(RNAi) animals displayed a weakly Uncoordinated phenotype. Vulval induction was scored in surviving progeny laid at least 6 hr postinjection (for the wild-type background), or in progeny laid 6–8 hr postinjection, before the lethal period began [for the let-60(gf) background]. In the wild-type background, sur-6 (RNAi) caused variable vulval defects, including defects in vulval induction (2° to 3° or hybrid fate transformation; 5/31 animals), VPC generation (absence of P5.p, P6.p, or P7.p, or failure of those cells to divide; 10/31 animals), and vulval lineage execution (failure of P5.pxx to complete last round of vulval division; 1/31 animals). The VPC generation defects were almost never seen in the let-60(n1046gf) background. In the wild-type background, PP2A-C RNAi also caused occasional defects in vulval induction (P5.p executed a “hybrid” lineage, generating only 3 vulval descendants and 1 nonvulval descendant; 1/23 animals), VPC generation (absence of P5.p; 1/23 animals), and vulval lineage execution (failure of P5.pxx to complete last round of vulval division; 1/23 animals). %Vul (and induction) are calculated only for animals in which P5.p, P6.p, and P7.p were present and divided at least once. %Muv (and induction) are calculated only for animals in which P(4-8).p were present and divided at least once.
We used the col-10 promoter to coexpress both sense and antisense RNA fragments in the hypodermis (see Material and Methods).