TABLE 2.
Rescue experiment
| Line | Obtained transgenic lines | Rescued | Not rescued | Rescuing ER stress of F1 |
|---|---|---|---|---|
| Phut-1::egfpa | 3 | 0 | 3 | − |
| PCR product of hut-1 | 3 | 3 | 0 | + |
| Phut-1::hut-1::egfpa | 2 | 2 | 0 | + |
| Peft-4::hut-1::Venusa | 5 | 5 | 0 | + |
| Peft-4::egfp::hut-1a | 5 | 3 | 2 | − |
| Peft-4::hUGTrel1 | 2 | 2 | 0 | − |
| Peft-4::schut1 | 3 | 0 | 3 | − |
| Peft-4::sphut1 | 4 | 0 | 4 | − |
| Peft-4::ZK896.9 | 2 | 0 | 2 | − |
| Peft-4::sqv-7 | 2 | 0 | 2 | − |
DNA was injected to 20–30 tm1435/+;zcIs4 hermaphrodite adults, and at least 50 F1 tm1435 heterozygotes, which produced tm1435 homozygous progeny strongly expressing Phsp-4::GFP, were screened. Obtained transgenic lines were cultured for 3 generations by blindly picking 8 individuals at each generation. Transgenic rescue was determined by the emergence of non-Scr tm1435 homozygous adults.
a
Fluorescence of Phsp-4::GFP can be easily distinguished from that of Phut-1::egfp, Phut-1::hut-1::egfp, Peft-4::hut-1::Venus, and Peft-4::egfp::hut-1 because their expression patterns are different.