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Adults were bred in spawning cages according to established protocols to obtain staged embryos (Westerfield 2000).
The bleached eggs were synchronized to L1 in M9 buffer at 20 °C for 16 h and transferred to IPTG (1 mM) containing NGM plate seeded with E. coli HT115 (DE3)/ L4440 and HT115 (DE3)/ L4440-G6PD at 20 °C for 3 days to obtain staged adults of mock and G6PD-knockdown C. elegans, respectively.
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Fertilized White Leghorn chicken eggs (E. Pavillard, Orny, Switzerland) were incubated at 38°C to obtain stage HH 3 4, stage HH 8, or stage HH 11 embryos [27].
It offers the possibility to obtain staging systems for other diseases, as we show here for Alzheimer's disease.
Fertilized chicken eggs obtained from a local breeder were incubated at 37 °C and 80%% relative humidity for the required time to obtain stages 17 18HH.
Embryos were collected and aged at 25° or 18° to obtain stages of interest, and standard protocols for fixing and staining were used.
Fertile Brown Bovan Gold hens' eggs (Henry Stewart, UK) were incubated for 1 16 hr to obtain stages X XIII (Eyal-Giladi and Kochav, 1976) and stage 4 (Hamburger and Hamilton, 1951) (HH).
In future work, it will be important to test this possibility and investigate the expression even closer to obtain stage-specific promoters which are powerful tools to meet different requirements.
We obtained stage grouping data in almost one in two cases.
To profile pNP expression, we mapped the obtained stage-specific Illumina paired-end reads to all pNP transcripts.
The renal tissue obtained was staged according to the WHO classification for lupus nephritis [ 30].
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