Fig. 6: NOTCH1 and NOTCH-2 ICD regulate similar transcriptomes in MCL. | Nature Communications

Fig. 6: NOTCH1 and NOTCH-2 ICD regulate similar transcriptomes in MCL.

From: Viral transduction of primary human lymphoma B cells reveals mechanisms of NOTCH-mediated immune escape

Fig. 6

a Box plots (1st to 3rd quartile, center = median) showing the expression levels of NOTCH1 and NOTCH2 genes in a series of CLL (n = 54) and MCL (n = 53) primary samples. The NOTCH1 or NOTCH2 mutational status of each sample is color labeled as indicated. RAM = robust multi-array average. b Unsupervised principal component analysis (PC2/3) of the RNAseq profiles of n = 3 paired MCL primary cases transduced with either an empty vector control, a NOTCH1ΔPEST or a NOTCH2ΔPEST expressing vector. A matrix of 40,490 transcripts was used to generate the unsupervised PCA. c Scatter plot showing the gene expression of the top 500 most variable genes in NOTCH1ΔPEST or NOTCH2ΔPEST expressing samples (n = 3). Pearson correlation of their expression in the NOTCH1ΔPEST and NOTCH2ΔPEST samples per pair was calculated. Each color corresponds to one of the three paired independent samples analyzed. Error bands indicate the 95% percentile of the mean. d Heatmap showing expression levels of genes differentially expressed in control vs. NOTCH expressing MCL samples (i.e., both NOTCH1ΔPEST and NOTCH2ΔPEST together). Gene expression levels are indicated as row z-scores. e CIITA immunoblot of Jeko-1 cells 5 days post transduction with NOTCH1ΔPEST or an empty vector control. Proteins were extracted following cell sorting of GFP-positive cells. One of three independent experiments is shown. f HLA-DR expression quantified by flow cytometry of NOTCH1ΔPEST transduced MCL cells (n = 4). HLA-DR expression was analyzed on GFP-positive cells 5 days post transduction. Each symbol refers to an individual patient sample. (p = 0.021). g PD-L1 expression quantified by flow cytometry of transduced primary MCL (n = 4) and Jeko-1 cells. PD-L1 expression was analyzed on GFP-positive cells 5 days post transduction (p = 0.003). Cohorts are shown as mean ± SEM. Statistical analysis was done by paired t-tests (f) and unpaired t-test (g). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 and not significant (ns) P > 0.05.

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