Extended Data Fig. 8: The translocation of PK6, PK7, and PK8 under glucose stimulation and depletion.
From: Nuclear-localized pyruvate kinases control phosphorylation of histone H3 on threonine 11
(a), (c), (e), and (h) Cellular sublocations of PK6-GFP (a), PK7-GFP (c), PK8-GFP (e), and GFP alone (h) in Arabidopsis protoplasts under glucose stimulation and glucose plus AZD8055 or Torin2. H3-RFP was used as a nuclear marker. GFP, H3-RFP, bright-field microscopy (differential interference contrast; DIC), and merged images are shown. Scale bars, 10 µm. GFP alone (h) was used as the negative control. GFP, green fluorescent protein. RFP, red fluorescent protein. (b), (d), (f), and (i) Quantitative confocal imaging of PK6-GFP (b), PK7-GFP (d), PK8-GFP (f), and GFP alone (i) in Arabidopsis protoplasts, determined by calculating the cytoplasm/nucleus (C/N) signal intensity ratio. Values shown are means ± SD; over 10 cells were scored for each treatment. Different letters mark statistically significant differences, as revealed using a one-way ANOVA (p < 0.01). (j) and (k) H3T11ph in Col-0 and PK6 levels in 35S:GFP-PK6 plants were measured during a darkness plus glucose and re-light treatment (j), and constituted long day plus glucose treatment (k) using a western blot. Molecular mass markers in kilodaltons (kDa) are indicated on the left in, and the bands are consistent with their expected sizes. H3 was used as a loading control. Experiments were repeated at least three times, and the results of representative experiments are shown. The source data underlying immunoblots are provided in Source Data File. (l) Relative transcription levels of PK6, PK7, and PK8 under long day photoperiod, darkness, and re-light conditions. Experiments were repeated at least three times. and the results of representative experiments are shown as means ± SE, n = 3. Ubiquitin 10 was used as internal control. The source data underlying the statistical analysis in (b), (d), (f), (i) and (l) are provided in the Source Data file.
