Trypanosomes lacking ZC3H30 are sensitive to stress.

Each panel shows cell numbers after application of a physical or chemical stress. Cells were incubated without selective drugs and were all in exponential growth at the start of the experiments. The DKO+ cell cultures included tetracycline for three days prior to, and for all of the time during, the experiments. Stresses were applied, then the cells were centrifuged and resuspended in fresh medium. In some experiments cells were diluted as necessary. Additional stresses are shown in S4A–S4C Fig. In individual experiments, the replicates for DKO and DKO+ were for independent cultures, usually of different clones. When experiments were repeated, 2 clones were usually used. (A) Bloodstream forms, 1 h at 42°C. This is a single experiment and individual replicates are shown. For a similar experiment at 42°C see S4A Fig. (B) Bloodstream forms, 1 h with 0.1 mM DTT. This is a single experiment and individual replicates are shown. (C) Procyclic forms, 1 h at 37°C. This is a single experiment with 3x WT, 3x DKO (each in triplicate), and 3x DKO+. Standard deviations are not visible beneath the data points. (D) Procyclic forms, 1 h at 41°C. This is a single experiment with 1x WT, 4x DKO, and 3x DKO+. (E). Procyclic forms, 1 h with 10 μM sodium arsenite. This is from two experiments. Experiment 1 had time points 24h and 48h, with 1x WT, 3x DKO, and 3x DKO+. Experiment 2 had time points 3h, 18h, 24h and 48h, with 3x WT, 6x DKO, and 5x DKO+. For a similar experiment with 20μM sodium arsenite see S4B Fig. (F) Procyclic forms, 1 h at 1% ethanol. This is a single experiment, with 1x WT, 3x DKO, and 3x DKO+. For a similar experiment with 2% ethanol see S4C Fig. (G) Procyclic forms, 3 h in PBS. This is from two experiments. Experiment 1 had 2x WT, 3x DKO, and 2x DKO+. Experiment 2 had 1x DKO and 2x DKO+. (H) Procyclic forms, 5 h in PBS. This is a single experiment and individual replicates are shown.