Fig. 6: c-Abl activation protected nerve cells from HAX-1 insufficiency-induced ROS accumulation and death.
From: Anti-apoptotic HAX-1 suppresses cell apoptosis by promoting c-Abl kinase-involved ROS clearance
A The MCF-7 scramble or HAX-1 knockdown cell line (MCF-7/siHAX-1) was treated with the indicated concentration of DPH for 12 h, and cell lysates were analyzed by immunoprecipitation and immunoblotted with indicated antibodies. The immunoprecipitates were normalized by c-Abl level. B MCF-7 scramble or HAX-1 knockdown cell lines (MCF-7/siHAX-1) with the indicated H2O2 (3 h) or DPH (4 h) treatment were subjected to flow cytometry analysis to evaluate ROS levels (left panel) and ratio of cell death (right panel). n.s., not significant; *p < 0.05, Student’s t test. C PMN-like cells were transfected with lentivirus-HAX-1-siRNA or lentivirus-Scramble-siRNA (MOI = 5), and HAX-1-siRNA cells were treated with DPH (10 μM, 12 h) or GSH (1 mM, 3 h). ROS levels and ratio of cell death of all the cells were measured after exposure to 1 mM H2O2 for 3 h (left panel) or for 8 h (right panel), respectively. *p < 0.05, **p < 0.01, Student’s t test. D Scramble cells, siRNA-HAX-1 SY5Y cells, and siRNA-HAX-1 SY5Y cells treated with nilotinib (5 μM, 18 h) or DPH (10 μM, 18 h) were subjected to 0.5 mM H2O2 treatment for 2 h, and the death rate was analyzed. n.s., not significant; *p < 0.05, **p < 0.01, Student’s t test. E WT or Hax-1 null mice were tail vein injected with DPH or vehicle daily for 30 days, and the striatum were subjected to terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. TUNEL-positive cells are marked in green, and nuclei are marked with DAPI. Right panel, quantification and statistical analysis of TUNEL-positive cells. **p < 0.01, Student’s t test.
