Journal: bioRxiv
Article Title: The CNS Microenvironment Promotes Leukemia Cell Survival by Disrupting Tumor Suppression and Cell Cycle Regulation in Pediatric T-cell Acute Lymphoblastic Leukemia
doi: 10.1101/2023.09.01.555887
Figure Lengend Snippet: A) RNA-sequencing analysis of normalized FPKM values showing decreased expression of SETD2 in CNS3 diagnosis patients compared to CNS1 diagnosis samples. (Significance calculated by two-tailed, unpaired Student’s t-test. *p<0.05). B) RNA-sequencing correlation analysis of SETD2 and TP53 (left graph) showing a negative correlation (r=-0.694, **p=<0.01) and MDM2 (right graph) showing a positive correlation (r=0.628, *p=<0.05) in CNS relapsed T-ALL patients. (Correlation was calculated using Pearson correlation coefficients, two-tailed with 95% confidence interval). C) String diagram showing protein-protein interactions directly and indirectly experimentally linking SETD2 with TP53, MDM2, BCLX, BCL2, NANOG and OCT4. D) mRNA expression levels (fold change versus empty vector control) of MDM2, TP53, MYCC, OCT4, NANOG, BCL2 and BCLX 24 hours after SETD2 overexpression by transfection in MOLT16 cells. E) mRNA expression levels of SETD2 overexpression in T.-ALL cell lines MOLT16, Jurkat, CCL-119 negatively correlates with BCL2 (r=0.-995, p=0.066), BCLX (r=-0.969), MDM2 (r=-979) and TP53 (r=-0.993, p=0.075). (Correlation was calculated using Pearson correlation coefficients, two-tailed with 95% confidence interval).
Article Snippet: DH5α competent cells (Invitrogen) were transformed by heat shock in 42°C water bath using SETD2 Human Tagged ORF Clone (Origene, RG224760) or pCMV6□AC□GFP Mammalian Expression Vector (Origene, PS100010) followed by overnight culture on LB agar plates (Sigma-Aldrich) containing 100μg/ml ampicillin (Sigma-Aldrich) at 37°C.
Techniques: RNA Sequencing Assay, Expressing, Two Tailed Test, Plasmid Preparation, Control, Over Expression, Transfection
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