4.2. Treatment of Auxenochlorella Pyrenoidosa by TOR Inhibitors

The A. pyrenoidosa cells was inoculated into a 50 mL BG11 liquid medium supplemented with different concentrations of TOR inhibitors (rapamycin, AZD8055, KU0063794, Torin1) and incubated at 28 °C, 2000 lux continuous light, and 180 rpm. The cell density at 680 nm optical density (OD680) was measured with a Microplate Reader (Biotek EpochTM2, Winooski, VT, USA) at 0, 2, 4 and 6 days.

To test whether A. pyrenoidosa cells were killed by high concentrations of AZD8055. A. pyrenoidosa cells were treated with 1, 5 and 10 μM AZD8055 for 4 days, and AZD8055 was removed from the medium, then the pellet was resuspended with BG11 and adjusted to the same OD value. Meanwhile, the removed supernatant containing different concentrations of AZD8055 was added into fresh A. pyrenoidosa cells. The phenotype was observed after culturing with or without AZD8055 for 4 days.