Figure 1.

Isolation of the mdh2-1 (Lb9G9) Mutant Impaired in TAG Breakdown during N Recovery.

(A) TAG content 2 d after N deprivation and 1 d after N resupply in mixotrophically grown cells.

(B) Visualization of LDs in cells upon N resupply (1 d) by staining with BODIPY.

(C) The insertion site of the cassette APHVIII in mdh2-1.

(D) RT-PCR analysis.

(E) Immunoblot analysis using anti-MDH2 antibodies.

(F) Cell volume determination.

Cells were first grown mixotrophically in N-deprived conditions. After N resupply, cells were kept in the dark in MM medium (1 d) then were stained with BODIPY, and pseudocolors were used: lipid droplet in green and chlorophyll in red. The housekeeping gene used for RT-PCR is RACK1. For immunoblots, samples were loaded at equal total protein amount and stained by Coomassie blue. RACK1, Receptor of activated protein C kinase 1. Values are the mean of biological replicates (i.e., independent shaking flask cultures; n = 5, sd). Biological replicates refer to cells that were grown in independent flasks. Asterisks indicate statistically significant changes compared with the parental line dw15 by paired-sample Student’s t test (*P ≤ 0.05 and **P ≤ 0.01).