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Fig. 13. Graphical summary. After establishing the xenograft mouse model, human leukemia cells from spleen, BM, and CNS were isolated and, together with cells from nine individual BCP ALL cell lines (NALM-6, REH, RS4;11, KOPN-8, UoCB6, RCH-ACV, MHH-CALL2, <t>EU3,</t> and HAL-01) and PBMCs derived from six human healthy donors, were subjected to miRNA isolation and cDNA synthesis. The expression levels of six standards (5sRNA, SNORD44, RNU6, RNU1A1, miR-103a-3p, and miR-532-5p) in all samples were analyzed by qPCR. Four different algorithms (geNorm, BestKeeper, Normfinder, ΔCT) were applied, identifying miR- 103a-3p and miR-532-5p as the most reliable references to be used in accurate qPCR normalization in miRNAs studies in BCP ALL xenografts. NOD/SCID/huALL: non-obese diabetic/severe combined immunodeficiency/ human acute lymphoblastic leukemia; BM: bone marrow; CNS: central nervous system; PBMC: peripheral blood mononuclear cell. Created in BioRender. Meyer, L. (2024) https://BioRender.com/z55z209
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Fig. 13. Graphical summary. After establishing the xenograft mouse model, human leukemia cells from spleen, BM, and CNS were isolated and, together with cells from nine individual BCP ALL cell lines (NALM-6, REH, RS4;11, KOPN-8, UoCB6, RCH-ACV, MHH-CALL2, EU3, and HAL-01) and PBMCs derived from six human healthy donors, were subjected to miRNA isolation and cDNA synthesis. The expression levels of six standards (5sRNA, SNORD44, RNU6, RNU1A1, miR-103a-3p, and miR-532-5p) in all samples were analyzed by qPCR. Four different algorithms (geNorm, BestKeeper, Normfinder, ΔCT) were applied, identifying miR- 103a-3p and miR-532-5p as the most reliable references to be used in accurate qPCR normalization in miRNAs studies in BCP ALL xenografts. NOD/SCID/huALL: non-obese diabetic/severe combined immunodeficiency/ human acute lymphoblastic leukemia; BM: bone marrow; CNS: central nervous system; PBMC: peripheral blood mononuclear cell. Created in BioRender. Meyer, L. (2024) https://BioRender.com/z55z209

Journal: Scientific reports

Article Title: Benchmarking miRNA reference genes in B-cell precursor acute lymphoblastic leukemia.

doi: 10.1038/s41598-024-77733-8

Figure Lengend Snippet: Fig. 13. Graphical summary. After establishing the xenograft mouse model, human leukemia cells from spleen, BM, and CNS were isolated and, together with cells from nine individual BCP ALL cell lines (NALM-6, REH, RS4;11, KOPN-8, UoCB6, RCH-ACV, MHH-CALL2, EU3, and HAL-01) and PBMCs derived from six human healthy donors, were subjected to miRNA isolation and cDNA synthesis. The expression levels of six standards (5sRNA, SNORD44, RNU6, RNU1A1, miR-103a-3p, and miR-532-5p) in all samples were analyzed by qPCR. Four different algorithms (geNorm, BestKeeper, Normfinder, ΔCT) were applied, identifying miR- 103a-3p and miR-532-5p as the most reliable references to be used in accurate qPCR normalization in miRNAs studies in BCP ALL xenografts. NOD/SCID/huALL: non-obese diabetic/severe combined immunodeficiency/ human acute lymphoblastic leukemia; BM: bone marrow; CNS: central nervous system; PBMC: peripheral blood mononuclear cell. Created in BioRender. Meyer, L. (2024) https://BioRender.com/z55z209

Article Snippet: Cell lines and cell culture The ALL cell lines NALM-6, REH, RS4;11, KOPN-8, RCH-ACV, MHH-CALL2, EU3, and HAL-01 were obtained from the German Collection of Microorganisms and Cell Cultures GmbH (DSMZ, Braunschweig, Germany).

Techniques: Isolation, Derivative Assay, cDNA Synthesis, Expressing