Fig. 5

Cells lacking Pkh are hypersensitive to oxidative stress. a β-Galactosidase activity of cells containing the STRE-LacZ reporter gene. SDP8 were transformed with the pGM18/17 plasmid, which was previously linearized at the URA3 gene marker by digestion with NotI, and treated as described for the Fig. 4a. β-Galactosidase activity was measured in non-treated (empty bars) or treated (filled bars) cells. Data are mean ± SEM from three independent clones. Treated cells were compared with untreated cells at each time point using un-paired t-test (n = 3; one-tailed P value; *P < 0.05). b Effect of partial depletion of Pkh on the sensitivity to oxidative agents. Wild type (WT) CML476 cells and the indicated mutants were grown, and four different (1:5) dilutions of the cultures containing the same number of cells were spotted on YPD plates containing 10 μg/ml doxycycline and the indicated concentrations of H₂O₂, menadione and diamide. Growth was monitored after 3 days. c Overexpression of YAP1 partially rescues the lethal effect of depletion of Pkh. MB002 and MB005 cells were transformed with the empty centromeric pRS316 or the same plasmid containing the promoter and coding region of YAP1. Serial dilutions of liquid cultures were spotted on YPD plates containing 50 μg/ml doxycycline (DOX) or the vehicle. Cell growth was recorded after 3 days