Growth curves and Northern blot of the TbINO1 conditional double knockout cell line.
A–D. Cells were washed with tetracycline-free HMI-9, and transferred to tetracycline-free media and counted daily. Growth curves are shown for wild-type cells in this medium (A), TbINO1 conditional double knockout with added tetracycline (B), TbINO1 conditional double knockout without tetracycline (C) and TbINO1 conditional double knockout without tetracycline, with added myo-inositol (100 mM) (D).
E. A Northern blot of total RNA from procyclic cells (lane 1), wild-type cells (lane 2), TbINO1 conditional double knockout without tetracycline on day 0 (lane 3), TbINO1 conditional double knockout without tetracycline on day 2 (lane 4) and TbINO1 conditional double knockout without tetracycline on day 12 (lane 5). The blot was either probed with TbINO1 ORF or β-tubulin, as detailed in Experimental procedures.