Extended Data Fig. 8. 14-3-3 protein is crucial for p300 accumulation in nucleus.
a, Localisation of phosphorylated p300 (p-p300) in complete and AA-depleted media using a p300 phospho-specific antibody. N = 3, n = about 30 cells analysed per condition per experiment. Scale bar, 5 μm. b, Interaction of His-tagged p300 with myc-tagged 14-3-3ζ and its increase in AA-depleted conditions. The expression levels following transfection are determined by using antibodies specific to the tagged protein. Blots are representative of three biologically independent experiments (N = 3). One-way ANOVA with post hoc Tukey test. c,d, Inhibition of interaction of p300 with 14-3-3ζ by 5 μM Comp. C. The expression levels following transfection are determined by using antibodies specific to the tagged protein. N = 3. One-way ANOVA with post hoc Tukey test. e, Inhibition of His-tagged p300 SA mutant binding with myc-tagged 14-3-3ζ. The expression levels following transfection are determined by using antibodies specific to the tagged protein. N = 4. One-way ANOVA with post hoc Tukey test. f, Remaining cytoplasmic p300 levels and mTORC1 activity in 14-3-3ζ knockdown cells in AA-depleted media for 1 h. N = 3. One-way ANOVA with post hoc Tukey test. g, Inhibition of p300 nuclear accumulation by depletion of AAs in 14-3-3ζ siRNA transfected HeLa cells. N = 3, about 30 cells scored per condition per experiment. 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.