Fig. 3. The effect of phospho-leucine-rich repeat kinase 2 (pLRRK2) targeting drugs including DNK72 in 0317, 448T glioma stem cells (GSCs) and orthotopic mouse models. (A, B) Representative western blot images of pLRRK2, LRRK2 in two GSCs with different concentrations of DNK72. The pLRRK2 level gradually decreased as the concentration of DNK72 increased. (C, D) Cell proliferation assays performed using two GSCs with different concentrations of DNK72. All error bars represent mean±SEM (n=3). **p<0.01, t-test. (E, F) Representative western blot images of pLRRK2, LRRK2 in two GSCs with different inhibitors of pLRRK2. pLRRK2 targeting drugs in a clinical trial for Parkinson’s disease treatment, PF-06447475, GNE7915, MLi-2, and LRRK2-IN-1, and we compared the effect of these drugs with DNK72 for the inhibition of pLRRK2 level. (G, H) Cell proliferation assays performed using two GSCs with different inhibitors of pLRRK2. All error bars represent mean±SEM (n=3). **p<0.01, t-test. The treatment of 1,000 nM of these compounds gave similar pLRRK2 inhibition capability (E, F), while DNK72 reduced cell proliferation slightly better than other compounds (G, H). (I) Kaplan–Meier survival plots for the orthotopic xenograft mouse model (log-rank test). The shLRRK2 treated orthotopic mouse models showed dramatically decrease of tumor size and increase of overall survivals. (J) H&E staining of the whole brains from mice implanted with 448 cells with different inhibitors of pLRRK2. Scale bars=1 mm.