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Journal: Cell Stem Cell
Article Title: Controlling genetic heterogeneity in gene-edited hematopoietic stem cells by single-cell expansion
doi: 10.1016/j.stem.2023.06.002
Figure Lengend Snippet: Single-cell cloning of gene-edited functional HSCs (A) Schematic showing the extracellular domain of CD45 with allele-specific antibody clones 104 and A20 and the epitope-defining amino acid. (B) Experimental setup of the single-cell editing and expansion experiment. (C) Left: fractions of CD201 + CD150 + KSL cells in single-cell-derived cultures 14 days after cloning (n = 261 clones). Right: Histogram of CD201 + CD150 + KSL cell frequency. Zoomed-in region shows clones with >10% CD201 + CD150 + KSL cells. (D and E) CD45.1 + donor PB chimerism (D) and lineage distribution (E ) in single recipients with long-term (LT) engraftment ≥5% and multilineage reconstitution (n = 8). Numbers over graphs in (D) represent percentage of CD201 + CD150 + KSL cells in the transplanted clone (%). (F) Linear correlation plots of CD201 + CD150 + KSL cell frequency and 16-week donor chimerism. Red dots indicate LT repopulating and multilineage clones. Pearson correlation. (G) CD45.1 + PB chimerism and lineage distribution in secondary recipients (n = 5). See also Figure S4 and Table S3 . Error bars represent SD.
Article Snippet: anti-mouse CD45.1-APC/Cy7 (A20) , Tonbo Biosciences , Cat#25-0453; RRID: AB_2621629.
Techniques: Cloning, Functional Assay, Clone Assay, Derivative Assay