Fig. 4.

CREBBP/EP300 mutations inhibited FBXW7 and activated the NOTCH signaling pathway. a Heatmap of genes associated with the NOTCH signaling pathway in CREBBP^mut/EP300^mut patients, as compared to CREBBP^wt/EP300^wt patients. b Normalized mRNA expression of FBXW7, HEY1 and HEY2 in tumor samples of diffuse large B-cell lymphoma (DLBCL) patients with or without CREBBP/EP300 mutations as revealed by RNA sequencing data. c Relative gene expression of FBXW7, HEY1, and HEY2 in CREBBP^R1392*, CREBBP^kd, EP300^H1377R, and EP300^kd DB cells, as compared to CREBBP^wt, EP300^wt or scramble DB cells by quantitative real-time PCR (RT-PCR). Data are presented as the mean ± SD (N = 3). d Protein expression of FBXW7 and NICD detected in vector, CREBBP^wt, CREBBP^R1392*, scramble, CREBBP^kd DB, SUDHL4, and LY10 cells, and in vector, EP300^wt, EP300^H1377R, scramble, EP300^kd DB, SUDHL4 and LY10 cells by western blot. The same lysates were used as in Fig. 3b. Tubulin was used as a loading control. The FBXW7/Tubulin and NICD/tubulin ratio were shown. e Occupancies of H3K27ac in the proximal promoter areas of FBXW7 in CREBBP^wt, CREBBP^R1392*, scramble, CREBBP^kd DB cells, and in EP300^wt, EP300^H1377R, scramble, EP300^kd DB cells by chromatin immunoprecipitation (ChIP) assay. Data are presented as the mean ± SD (N = 3). f Expression of NICD in CREBBP^R1392* and CREBBP^kd DB cells, as well as EP300^H1377R and EP300^kd DB cells with or without reintroduction of FBXW7 protein by western blot