Notch inhibition decreases DNTs. (A) Response of DNTs in peripheral blood to GSI in 6 treated and 13 contorl CBA-lprcg mice. Retro-orbital bleeds were performed every 2 weeks to assess absolute DNTs/mm3, comparing mice treated with DAPT and vehicle control (absolute DNTs: WBC in mm3 × % CD3+/TCRα/β+/CD4−/CD8− cells) Serial absolute DNT measurements were normalized to the untreated baseline for each mouse. Treated mice showed a statistically significant decrease in absolute DNTs compared with controls by 6 weeks of treatment. Bars represent normalized mean absolute double negative T-cell counts from mice at each time point, and error bars represent SEM. (B) Average number of DNTs in lymph nodes and spleens (defined as total cell count × % CD3+/TCRα/β+/CD4−/CD8− cells), comparing GSI-treated and control animals, showing a statistically significant decrease in both. Error bars represent SEM.