Figure 1
From: Snf1/AMPK is involved in the mitotic spindle alignment in Saccharomyces cerevisiae
(a) A Snf1-HA expressing strain was grown in synthetic medium containing 2% glucose until exponential phase, then arrested and released from HU block as described in Materials and Methods. Samples were taken at different time points (0, 45, 60, 75, 90, 105 minutes) and analysed by western blot with anti-pT172-AMPK and anti-HA antibodies. Budding index (BI%) of cells at each time point and densitometric analysis of the pSnf1/tot ratio are shown. Cells expressing Snf1-T210A-HA and Snf1-HA cells shifted to 0.05% glucose for 10 min were used as negative and positive control, respectively. Controls were run together with samples and cropped as indicated by vertical lines. (b–d) Immunostaining for pT210-Snf1 using anti-pT172-AMPK antibodies and DAPI staining of DNA. bar: 5 µm. (e) Example of asynchronous snf1Δ cells with misaligned metaphase spindle. Spindle morphology (tubulin) and DAPI are shown. bar: 5 µm.
