Extended Data Fig. 3. p300 nuclear transport under glucose starvation.
a, p300 nucleocytoplasmic shuttling in AAs or Glc-depleted media for 1 h using fractionation. Blots are representative of three biologically independent experiments (N = 3). One sample t-test. b, p300 nucleocytoplasmic shuttling in Glc-depleted media for 1 h, and then re-addition of Glc to the media for 1 h. N = 3, about 50 cells scored per condition per experiment. One-way ANOVA with post hoc Tukey test. Scale bar, 5 μm. c, Remaining mTORC1 activity in His-tagged p300 dNLS or S89A expressing p300 KO cells in Glc-depleted media for 1 h. The expression levels following transfection are depicted by the quantification of the His-tag. N = 4. One-way ANOVA with post hoc Tukey test. d, Rescue of autophagy activation in raptor knockdown cells by reconstituting with wild-type (WT) raptor, not KR mutant (K1097R). N = 4. One-way ANOVA with post hoc Tukey test. e, Autophagic flux with HeLa cells stably expressing mRFP-GFP-LC3 reporter. Representative confocal z-stack images (left panel) and total number of mRFP/GFP dots (autophagosomes) and mRFP-only dots (autolysosomes). N = 4; 60-80 cells analysed per condition per experiment. Scale bar, 5 μm. One-way ANOVA with post hoc Tukey test. f, Increased LC3-II levels by expressing raptor KR mutant in p300 KO cells reconstituted with p300 WT or dNLS. N = 4. One-way ANOVA with post hoc Tukey test. g, Glucose uptake assay in p300 KO cells expressing His-tagged WT, dNLS or S89A. Insulin (1 µM) was used to stimulate glucose uptake into cells. N = 4 in each group. One-way ANOVA with post hoc Tukey test. Data are presented as mean values +/- s.d. unless otherwise specified. Source numerical data and unprocessed blots are available in source data.