Fig. 4. Rapamycin effects on different insensitive cells.

A Fold-change in the proliferation of untransduced cells treated with 109 nM rapamycin (TT) or not (UT) and weaned after 48 h measured by Trypan blue counting. Counting was performed each day for 168 h. Mean ± SD of three independent experiments. B Left, experimental design for the proliferation analysis by carboxyfluorescein succinimidyl ester (CFSE) labeling during and after rapamycin treatment; right, representative histograms of CFSE labeling of T cells treated with rapamycin and weaned after 48 h. CD3⁺/CFSE⁺ cells were analyzed by flow cytometry 96 h after rapamycin treatment and 192 h after weaning. C The impact of rapamycin on CD19 (left) and CAR IL-1RAP (right) expression in iCasp9-expressing genetically modified T cells. Cells were treated for 24 h with 109 nM rapamycin (TT, treated) or not (UT, untreated) and then weaned. CD3⁺/CD19⁺/CAR IL-1RAP⁺ cells were analyzed 24 h after treatment began and 72 h and 96 h after weaning. **P < 0.01; with Two-way Anova test.