Fig. 4

NFκB signaling mediated TNFα-induced enhanced CD7⁺ HTLP generation. A The predicted regulatory pathway network by IPA based on differential gene expression by CD34⁻CD7⁺ cells generated in the presence or absence of TNFα. IPA was performed after RNA sequencing of sorted CB or mPB HSPC-derived CD34⁺CD7⁺ and CD34⁻CD7⁺ progenitors after 7 days of DL-4 culture. B A representative FACS histogram showing the phosphorylation of NFκB (left panel) and corresponding MFIs (right panel) at the indicated time points after treatment with TNFα during HTLP culture (mean ± SEM, n = 2). The p values were calculated by an unpaired two-tailed t test: *p ≤ 0.05; **p ≤ 0.01. C A representative FACS plot of the phenotype of 7-day HTLP cultures of mPB HSPCs in the presence (100 ng/ml) or absence of TNFα or in the presence of TNFα and piceatannol (25 µM) (an NFkB inhibitor). Graphs showing the mean frequencies (D) and numbers (E) of CD34⁺CD7⁺ progenitors (in black) and CD34⁻CD7⁺ progenitors (in light gray) after 7 days of HTLP culture of mPB HSPCs in the presence (100 ng/ml) or absence of TNFα or in the presence of TNFα and piceatannol (25 µM) (an NFkB inhibitor) (mean ± SEM, n = 3). The p values were calculated by one-way ANOVA: ***p ≤ 0.001