For qRT PCR, an equal volume (100 μl for plasma and 200 μl for cell-culture media) of sample was processed in RNA isolation for each subject.
For irrigation, an equal volume (0.5 L per day) of freshwater was applied to each pot in the morning.
For precipitation, an equal volume of TCA (20% wt/vol) was added to a final concentration of 10% (wt/vol) and incubated at 4°C for at least one hour until overnight.
For this, an equal volume of ice-cold 5% TCA was mixed with aliquots of 2 µg of each Eu-labeled protein diluted in 200 µl of distillated water followed by centrifugation at 3,000 g for 10 minutes, removal of the supernatant, and resuspension of the pellet in Delfia enhancing solution (Perkin-Elmer) by vortexing.
Briefly, worms were either cut (for immunostainings) or not (for PI and phalloidin stainings), placed in PBST with 3.2% paraformaldehyde for fixation, with an equal volume of heptane, under rotation for 10 minutes.
For Western blotting, an equal volume of 2x SDS-PAGE buffer was added to the periplasmic and soluble protein fractions and then boiled for 15 min at 100°C.
After incubation on ice for 10 min, an equal volume of neutralization solution (400 mM HCl, 600 mM Tris HCl) was added.
After centrifuging at 10,000 g for 10 min, an equal volume of 1M NaOH was added to the supernatant and absorbance was measured at 450 nm in both samples and controls.
For vehicle controls, an equal volume of 100% ethanol was added in the same manner.
Briefly, enriched T cells were resuspended in PBS for staining with an equal volume of 0.8 μM CFSE.
