Fig 1: UNC119 inhibition reduces proliferation in T-ALL cell lines and PDX samples.(A) Western blot showing the expression of pLCK Y394 in healthy CD8+ T cells (OTI) compared with Jurkat, MOLT4, and CCRF T-ALL cell lines relative to cofilin as a loading control. (B) Western blot showing the expression of UNC119 in Jurkat, MOLT4, and CCRF cell lines relative to actin as the internal loading control. (C) Confocal imaging of endogenous pLCK in T-ALL cells treated with either DMSO or Unc119i. White arrows on images indicate intracellular LCK stores. Representative images are from two independent experiments. Scale bars represent 5 μm. (D) Graphs showing the median fluorescence intensity of pZAP70 Y319 in human T-ALL cell lines with either DMSO, 5 μm Unc119i, or 2 μm LCKi (MedChemExpress—HY12072). Data shown are from two independent experiments, where each dot represents technical replicates from two experiments. Statistical significance was determined with one-way ANOVA on Prism GraphPad. (E) Proliferation curves of Jurkat, MOLT4, and CCRF cells with increasing concentrations of Unc119i and a fixed concentration of LCKi at 2 μm. Each condition and time point was plated in triplicate, each data point representing the average of three technical replicates. Error bars represent the standard error of the mean. One dose–response experiment was conducted per T-ALL cell line. (F) Graphs showing the fold change in proliferation at endpoint of the same three human T-ALL cell lines. Four independent experiments were conducted. Each condition was plated in triplicate, and fold change was calculated by dividing the average cell concentration at 96 h by that at 0 h. Statistical significance was determined with one-way ANOVA on Prism GraphPad. (G) Graph showing the percentage of Annexin V+ Zombie NIR+ apoptotic T-ALL cells with increasing concentrations of Unc119 inhibitor. (H) Proliferation curves of four different PDX-derived human T-ALL cells in response to Unc119i and LCKi treatments. Each dot is the average of triplicate plating for each PDX sample. All error bars represent the standard error of the mean.Source data are available for this figure.