Figure 7.
MYC-induced translation impairs PLAG1-S-mediated stemness in human HSPCs. (A) Up- (red) or down- (blue) regulation of MYC ribosome biogenesis targets in PLAG1-SOE HSPCs. (B) Schematic of PLAG1-S and c-MYC overexpression lentivectors. (C) Representative sorting gates for dual-overexpression of PLAG1-S and c-MYC or controls in Lin−CD34+ cells. (D) CD34 (n = 4) and (E) CD33 (n = 3) positivity in BFP+GFP+ HSPCs over 4 and 7 days of ex vivo culture. (F) Cell size determined by flow cytometric MFI of FSC-H (n = 3-4) in BFP+GFP+ HSPC cultures on day 4 and 7. (G) OP-Puro incorporation by BFP+GFP+ HSPCs on day 4 of ex vivo culture (n = 4). Data are presented as average ± SEM unless otherwise indicated. Each point represents an individual CB unit. ***P < .005, **P < .01, *P < .05. n.s., not significant. See also supplemental Figure 7.

MYC-induced translation impairs PLAG1-S-mediated stemness in human HSPCs. (A) Up- (red) or down- (blue) regulation of MYC ribosome biogenesis targets in PLAG1-SOE HSPCs. (B) Schematic of PLAG1-S and c-MYC overexpression lentivectors. (C) Representative sorting gates for dual-overexpression of PLAG1-S and c-MYC or controls in LinCD34+ cells. (D) CD34 (n = 4) and (E) CD33 (n = 3) positivity in BFP+GFP+ HSPCs over 4 and 7 days of ex vivo culture. (F) Cell size determined by flow cytometric MFI of FSC-H (n = 3-4) in BFP+GFP+ HSPC cultures on day 4 and 7. (G) OP-Puro incorporation by BFP+GFP+ HSPCs on day 4 of ex vivo culture (n = 4). Data are presented as average ± SEM unless otherwise indicated. Each point represents an individual CB unit. ***P < .005, **P < .01, *P < .05. n.s., not significant. See also supplemental Figure 7.

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