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. 2017 May 31;114(24):6346–6351. doi: 10.1073/pnas.1617799114

Fig. 1.

Fig. 1.

C. albicans PHO84 is required for rapamycin tolerance, growth during Pi starvation, normal TORC1 activity and hyphal morphogenesis, and Pi homeostasis. (A) Cell dilutions of WT and a mutant series in PHO84 pinned onto YPD with vehicle or 12 ng/mL rapamycin. PHO84+/+, JKC915; pho84−/+, JKC1583; pho84−/−, JKC1450; pho84−/−/+, JKC1588. (B) Cells as in A pinned onto YNB with 0 or 11 mM Pi. (C) Separate Western blots of the same samples for P-S6, total Rps6, and tubulin of WT (+/+; JKC915), pho84 null (−/−; JKC1450), and PHO84 reintegrant (−/−/+; JKC1588) cells grown in YNB with 0, 0.22, or 11 mM KH2PO4 for 90 min. Dens, densitometric ratio of P-S6 vs. tubulin signal. (D) Strains as in A spotted at equidistant points around agar plates with spot edges imaged. Compare with SI Appendix, Fig. S2A. (Scale bar: 1 mm.) (E) S. cerevisiae and C. albicans WT and pho84 null cells grown in SC medium with 0.22 mM (low Pi) or 11 mM Pi (high Pi) overnight assayed for free and total Pi; pho85 null cells as controls, which hyperaccumulate Pi. Ca+/+ (C. albicans PHO84/PHO84 PHO85/PHO85), JKC915; Capho84−/−, JKC1450; Capho85−/−, CaLC1919 grown in 20 μg/mL doxycycline overnight; Sc+ (S. cerevisiae PHO84 PHO85), BY4741; Scpho84−, EY2960; Scpho85−, from ref. 37. Error bars: SDs of three technical replicates. *P < 0.01; **P < 0.05.