LNK inhibition enhances clonogenic potential of FA-like HSPCs in the presence of various DNA stressors. (A) Experimental scheme of lentiviral transduction followed by CFC. CB-derived CD34⁺ cells were first transduced with lentivirus-expressing GFP and either shLuc or shFANCD2 to establish FA-like HSPCs. Subsequently, the cells were transduced with lentivirus-expressing mCherry and either shLuc or shLNK #1, and 48 hours post–second transduction, GFP⁺mCherry⁺ cells were sorted and either directly plated in methylcellulose CFC assay with indicated concentrations of MMC or HU or were pretreated for 4 hours with acetaldehyde (Ace) and then plated in methylcellulose. Vehicle-treated cells were used as control. (B) CFU numbers were quantified after 12 to 14 days and a representative experiment is shown. (C-E) Percentages of CFUs in indicated drugs relative to vehicle-treated controls are shown. Each symbol represents an individual plate; bars indicate mean values; error bars indicate plus or minus SD. *P < .05; **P < .01; ***P < .001 as determined by 2-way ANOVA followed by Tukey’s multiple comparisons. For simplicity, in panels C-E, multiple comparisons for shLuc/shLNK are not shown.