Extended Data Fig. 6: Regulation of cyclin D1 transcription requires p300/CBP.
From: Cytosolic pH regulates proliferation and tumour growth by promoting expression of cyclin D1
(a, b) Reduced activity of p300/CBP decreases cyclin D1 expression and acetylation of histone H3 at K27. (a) Cells were subjected to shRNA against the indicated transcription factors or co-activators and histone acetylation at H3-K27 was determined by western blotting. (b) Cells were grown as in Fig. 3b and cyclin D1 mRNA levels were determined by 4 hours after stimulation with the indicated conditions by qPCR (mean ± S.E.M. of 3 independent experiments, two-sided t-Test). (c) Cells were grown as in Fig. 3a and the efficiency of shRNA mediated knock-down of p300 and CBP and cyclin D1 protein levels were determined by western blotting. (d) DMA treatment and p300/CBP inhibition does not generally affect histone acetylation. Cells were grown as in Fig. 3c and acetylation of histone H3 at K9 was determined by western blotting. (e–g) Starvation, inhibition of NHE activity and p300/CBP activity trigger a similar gene expression program. Cells were grown as in Fig. 1h and gene expression was determined by RNAseq. Volcano plots of differentially expressed genes upon stimulation with FCS glucose relative to (e) starvation, (f) DMA treatment and (g) C646 treatment is shown. Co-regulated genes under all conditions are indicated in red. (h) Venn diagram showing number of significantly higher expressed genes under the indicated conditions. For all western blots in this figure, blots representative of three independent experiments are shown.
