Fig. 3.

Yap1801 and Yap1802 proteins are upregulated during glucose starvation. (A) Immunoblot analysis of endogenously tagged Yap1801–mGFP and Yap1802–mGFP in wild-type cells grown in glucose and raffinose media. Anti-CPY antibody was used as a loading control. (B) Histogram showing average Yap1801–mGFP and Yap1802–mGFP intensity, normalized to intensity in glucose medium. Mean±s.d. of n=4. (C) Wild-type cells expressing Yap1801–mGFP and Yap1802–mCherry at mid-log phase were imaged by Airyscan confocal and Slimfield microscopy. For Slimfield, bright foci are indicated with white arrows, and the integrated density for diving cells in each frame is shown in yellow. These data represent the first of 200 frames, acquired every 5 ms, with representative full datasets shown in Movie 3. (D) Numbers of Yap1801 and Yap1802 molecules per cell (>20 cells) were estimated using autofluorescence- and background-corrected integrated density values obtained using ImageJ software. Mean±s.d. is shown. (E) Airyscan confocal colocalisation of Mup1–GFP with Yap1801–mCherry (upper panels) and Yap1802–mCherry (lower panels) in glucose and raffinose media. Yellow boxes denote zoomed-in regions shown in the panels to the right. *P<0.05 (unpaired Holm–Sidak t-test). Scale bars: 5 µm (white), 1 µm (yellow).