4.5. Cell Treatments

Exponentially-growing cells (five days) were treated with purified HYTLO1 (0.6 μM). The dose applied to cells was chosen on the basis of previous work concerning in vivo bioassays of physiological effects of HYTLO1 on plants [20]. In some experiments cells were treated with crude preparations of Myc factors or Nod factors. The final dose applied to cells corresponded to 10-fold concentrated AM fungal exudates and 1 µM Nod factors. In this latter case, suspension-cultured cells were grown in low nitrogen medium (containing 5 mM KNO₃) for two days prior to the onset of the Ca²⁺ measurement assays.