Fig. 4

Increased caspase-3 activation in PD patient-derived mDANs is reversed by treatment with NPT100-18A. (A) NPT100-18A- and DMSO-treated iPSC-derived mDANs (TUBB3⁺/TH⁺) from PD patients and controls were stained for cleaved caspase-3 (cCasp3) to evaluate rates of early neuronal cell death. Representative images used for quantification shown in (C). Scale bar 50 μm. (B) Immunocytochemistry quantification. mDANs from two PD patients (P) (with two different iPSC-clones for each patient) and three control individuals (with one iPSC clone for control [C] 1 and 2 and two clones for C3). No significant difference in pooled rates of TH/TUBB3-double positive neurons was observed between DMSO- and NPT100-18A-treated patient and control cultures, respectively. Control: two-tailed nested t-test P = 0.77; mean DMSO = 22.8%, mean NPT100-18A = 22.3%. Patient: two-tailed nested t-test P = 0.94; mean DMSO = 20.5%, mean NPT100-18A = 20.4%. (C) Treatment with 10 nM NPT100-18A significantly reduced rates of cCasp3-positive mDANs compared to vehicle condition in all patient-derived but not in control mDAN lines. Control: two-tailed nested t-test P = 0.86; mean DMSO = 3.7%, mean NPT100-18A = 3.9%. Patient: two-tailed nested t-test P = 0.004; mean DMSO = 14.4%, mean NPT100-18A = 4.2%; **P = 0.004. Values are shown as mean ± SD of three independent differentiations. Scale bar 50 μm