Journal: bioRxiv

Article Title: Proteasome-dependent degradation and nucleus–vacuole junctions sustain proteostasis during acute glucose starvation

doi: 10.64898/2026.04.22.720209

Figure Lengend Snippet: A) Representative immunoblots showing turnover of mNeonGreen-tGnd1-HA following 90 min of acute starvation (0.02% glucose) or in glucose-replete control medium (2% glucose). Protein stability was monitored by cycloheximide chase assay (100 µg/mL) at the indicated time points. Total protein loading was visualized and normalized using Stain-Free technology. Graphs represent mNG-tGnd1-HA protein levels as a percentage of the protein present at the time point 0 min. Data are presented as average ± SD from 2 independent experiments. B) Representative confocal fluorescence microscopy images logarithmically growing S. cerevisiae expressing mNeonGreen-tGnd1-HA, which were cultured in glucose-replete (2%) or glucose-deprived (0.02%) media for 90 min, followed by CHX chase (0 and 60 min) prior to fixation and imaging. Images represent maximum intensity projections of Z-stacks. Scale bar, 10 µ m. The bar chart shows the percentage of cells in the culture that contain inclusions, along with the distribution of cells harboring one and two or more inclusions. Data are presented as average ± SD from 2 independent experiments. C) Representative immunoblots showing turnover of mNeonGreen-stGnd1-HA after 90 min acute starvation (0.02% glucose) or control medium (2% glucose), assessed by cycloheximide chase assay (100 µg/mL) at indicated times (0, 60 min). Total protein loading was visualized using Stain-Free technology. Graphs represent mNG-stGnd1-protein levels as a percentage of the protein present at the time point 0 min, with average values and standard deviation (n = 2). D) Representative confocal fluorescence microscopy images of log-phase yeast cells expressing mNeonGreen-stGnd1-HA under glucose-rich YPD (2%) or glucose-deprived (0.02%) conditions for 90 min. Images represent maximum intensity projections of Z-stacks. Scale bar, 10 µ m.

Article Snippet: To investigate the cellular response to acute glucose starvation, yeast Saccharomyces cerevisiae cultures were grown in rich YPD medium (1% (w/v) yeast extract, 2% (w/v) peptone, and 2% (w/v) D-glucose; Formedium Ltd.).

Techniques: Western Blot, Control, Staining, Fluorescence, Microscopy, Expressing, Cell Culture, Imaging, Standard Deviation

Journal: bioRxiv

Article Title: Proteasome-dependent degradation and nucleus–vacuole junctions sustain proteostasis during acute glucose starvation

doi: 10.64898/2026.04.22.720209

Figure Lengend Snippet: A) Representative confocal fluorescence microscopy images of log-phase yeast cells expressing HA-mNG-NBD2* under glucose-rich YPD (2%) or glucose-deprived (0.02%) conditions for 90 min. Images represent maximum intensity projections of Z-stacks. Scale bar, 10 µ m. The bar chart depicts the proportion of cells in the culture that contain inclusions. Data are expressed as mean ± SD from two independent experiments. B) Representative immunoblots showing turnover of the model misfolded protein HA-mNG-NBD2*. Log-phase yeast cells were subjected to 90 min acute glucose starvation (0.02% glucose) or control 2% glucose YPD medium, followed by cycloheximide chase assay (100 µ g/mL) at the indicated time points. Total protein loading was visualized using Stain-Free technology.

Article Snippet: To investigate the cellular response to acute glucose starvation, yeast Saccharomyces cerevisiae cultures were grown in rich YPD medium (1% (w/v) yeast extract, 2% (w/v) peptone, and 2% (w/v) D-glucose; Formedium Ltd.).

Techniques: Fluorescence, Microscopy, Expressing, Western Blot, Control, Staining