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The ∆VIM mutant is deleted for amino acids 622–625. b, Serial tenfold dilutions of wild-type and vms1Δ cells transformed with the indicated vms1 alleles were spotted on YPD plates containing 10 or 25 ng ml−1 cycloheximide and incubated at 30 °C for 3 days.

Hadjiolova, K. V., Nicoloso, M., Mazan, S., Hadjiolov, A. A. & Bachellerie, J. P. Alternative pre-rRNA processing pathways in human cells and their alteration by cycloheximide inhibition of protein synthesis.

The cells were then either transfected with an empty p3×-Flag vector or Flag-tagged AIMP2 vector and grown for another 48 h before the addition of cycloheximide.

Immediately before sample collection, cultured mammalian cells were incubated with media containing 100 µgml cycloheximide for 10min at 37°C to stop translation elongation.

Soon after fertilization the jelly membranes of X. laevis embryos were chemically removed, and at the desired stages, embryos were flash frozen in lysis buffer without cycloheximide pre-treatment.