Figure 4.
Expression of DNMT3A mutations in a cytotoxic CD8+ PTCL cell line. (A, top) Representative Western blot of MYC-tagged DNMT3A. (A, bottom) Representative histogram of 5mC (MFI) in indicated T8ML1 cells. (B) Box plots of the log2 ratio (observed/expected) for 5mC peaks within the indicated genomic regions. (C) Pooled metagene plot for MeDIP-Seq profiles of DNMT3A-MT (R882H, V716D, and Q886Stop) and DNMT3A-WT (WT, EV) cell lines. Lines represent sample-type average log2 (peaks per bp per gene) for indicated regions, and shading represents standard error among samples. (D) Heatmap of concordant differentially expressed and methylated genes. (E) Supervised heatmap of representative differentially expressed and methylated genes. (F) UCSC genome browser visualization of MeDIP-Seq peaks in DNMT3A-MT samples and DNMT3A-WT samples for the listed genes. Gene diagrams represent the position of TSS relative to displayed genomic regions. (G) Normalized cell growth for T8ML1 cells with indicated DNMT3A alterations. Symbols represent mean ± standard error, n = 3 independent experiments. (H-J) Representative histograms of (H) DNMT3A expression, (I) phosphorylation status of LCK (pY505) and SLP-76 (pY128) under normal culture conditions, and (J) expression of T-cell development transcription factors. Representative histograms and blots are taken from n = 3 independent experiments. TSS, transcriptional start site; TTS, transcriptional termination site; MT, mutant; EV, empty vector.

Expression of DNMT3A mutations in a cytotoxic CD8+ PTCL cell line. (A, top) Representative Western blot of MYC-tagged DNMT3A. (A, bottom) Representative histogram of 5mC (MFI) in indicated T8ML1 cells. (B) Box plots of the log2 ratio (observed/expected) for 5mC peaks within the indicated genomic regions. (C) Pooled metagene plot for MeDIP-Seq profiles of DNMT3A-MT (R882H, V716D, and Q886Stop) and DNMT3A-WT (WT, EV) cell lines. Lines represent sample-type average log2 (peaks per bp per gene) for indicated regions, and shading represents standard error among samples. (D) Heatmap of concordant differentially expressed and methylated genes. (E) Supervised heatmap of representative differentially expressed and methylated genes. (F) UCSC genome browser visualization of MeDIP-Seq peaks in DNMT3A-MT samples and DNMT3A-WT samples for the listed genes. Gene diagrams represent the position of TSS relative to displayed genomic regions. (G) Normalized cell growth for T8ML1 cells with indicated DNMT3A alterations. Symbols represent mean ± standard error, n = 3 independent experiments. (H-J) Representative histograms of (H) DNMT3A expression, (I) phosphorylation status of LCK (pY505) and SLP-76 (pY128) under normal culture conditions, and (J) expression of T-cell development transcription factors. Representative histograms and blots are taken from n = 3 independent experiments. TSS, transcriptional start site; TTS, transcriptional termination site; MT, mutant; EV, empty vector.

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