Figure 7.
Cells without 4EIP cannot differentiate into growing procyclic forms. (A) Expression of various proteins in EATRO1125 cells cultured to high density for 60 h, then treated with cis-aconitate (CA) and transferred to 27°C. WT SL is long slender bloodstream forms; KO: no 4EIP; K+: knockout line with inducible 4EIP-myc and tetracycline; ΔN—knockout line with inducible lambdaN-ΔN-4EIP-myc, with tetracycline. In this experiment, full-length 4EIP-myc was not detected at time = 0. (B) Cell counts for cells treated as in (A) (to the end of day 1), then transferred to procyclic medium. (C) Expression of various proteins in cells cultured to 1 × 106/ml, then treated with cis-aconitate and transferred to 27°C. Replicates are in Supplementary Figure S8C. (D) Cell counts for cells from (C) transferred to procyclic medium on day 1.