Fig 5. C. albicans adapts to long term Ypd1 loss by lowering Hog1 activity.

(A) ypd1Δ cells become morphologically similar to wild-type cells over time. Micrographs of exponentially growing Wt (JC21) and ypd1Δ (JC2001) cells taken from rich media plates after 1 or 13 days. (B) ypd1Δ cells gradually accumulate stress phenotypes characteristic of wild-type cells. Approximately 10⁴ cells, and 10-fold dilutions thereof, of exponentially growing Wt, hog1Δ (JC50) and ypd1Δ cells taken from rich media plates after 1 or 13 days were spotted onto plates containing; NaAsO₂ (1.5 mM), calcofluor white (CFW 30 μg/ml) and NaCl (0.5 M). Plates were incubated at 30°C for 24 hrs. (C) Hog1 phosphorylation is not sustained in ypd1Δ cells over time. Western blot analysis of whole cell extracts isolated from exponentially growing Wt and ypd1Δ cells taken from rich media plates after the number of days indicated. Blots were probed for phosphorylated Hog1 (Hog1-P), stripped and reprobed for total Hog1 (Hog1). (D) PTP2, PTP3, and GPD2 expression is not sustained in ypd1Δ cells. Northern blot analysis of the indicated genes in exponentially growing Wt and ypd1Δ cells taken from plates after the number of days indicated. ACT1 was used as a loading control.