Figure 3.

Progranulin depletion induces caspase-independent apoptosis. a) Kasumi-1 cells were cultured with anti progranulin antibody (200 μg/ml) or control antibody for the indicated hours. Whole cell lysate was collected and expression level of canonical apoptosis related proteins was analyzed by western blotting. b) Expression level of apoptosis related proteins was normalized to that of β-actin. Error bars indicate the SD from mean; n = 4. (∗P < 0.05, ∗∗P < 0.01; two-tailed Student's t test) c) Kasumi-1 cells transfected progranulin-specific or control siRNA were cultured for 48 h and cell apoptosis was analyzed by using Annexin V & Dead Cell Kit. d) Progranulin specific siRNA or control siRNA transfected Kasumi-1 cells were cultured with or without Z-VAD-FMK (50 μM) for 48 h. Whole cell lysate was collected and PARP cleavage was analyzed by western blotting. e) Progranulin specific siRNA or control siRNA transfected Kasumi-1 cells were cultured with or without Z-VAD-FMK (50 μM) and proliferation of treated cells was determined by MTT assay. Error bars indicate the SD from mean; n = 3. (∗∗∗P < 0.001 between Ctrl and PGRN KD. †P < 0.05, ††P < 0.01 between Ctrl and Ctrl + Z-VAD-FMK. ‡‡P < 0.01 between PGRN KD and PGRN KD + Z-VAD-FMK; Tukey's test).