Fig. 3

SCAMP5 knockdown induces apoptosis. A PC12 cells were transfected with indicated siRNAs for 72 h and protein levels were analyzed by Western blot. Lysates were probed with indicated antibodies. The primary antibody was anti-α-synuclein. GAPDH served as a loading control. B Quantitative comparison of α-synuclein in PC12 cells. Scamp5 knockdown in PC12 cells resulted in α-synuclein accumulation. C Measurement of α-synuclein oligomers in PC12 cell by ELISA assay. D, E PC12 cells were transfected with the indicated siRNAs for 72 h, followed by flow cytometry analysis after staining with Propidium iodide and annexin V FITC. D The dot plot shows apoptosis and necrotic in control and treated cells. CON: control, UL: death cells, UR: late apoptosis cells, LR: early apoptosis cells, LL: normal cells. The red line indicates the position of the FITC line on the x-axis. E Quantitative measurement of the apoptosis cell. Data represent average population of representative experiments. F–I PC12 cells were transfected with indicated siRNAs for 72 h and protein levels were analyzed by Western blot. F, H Lysates were probed with indicated antibodies. The primary antibody was anti-Mcl-1, anti-Bcl2, anti-Bad, anti-caspase3, and anti-cleaved-caspase3. G, I Quantitative comparison of apoptosis-related factors in PC12 cells. Tubulin Beta 5 Class I (Tubulin5) served as a loading control. Data in panel B, E, G and I are presented as mean ± SEM. n ≥ 3 independent experiments. Significance levels are denoted as follows: ***P < 0.001SCAMP5 knockdown induces apoptosis. A PC12 cells were transfected with indicated siRNAs for 72 h and protein levels were analyzed by Western blot. Lysates were probed with indicated antibodies. The primary antibody was anti-α-synuclein. GAPDH served as a loading control. B Quantitative comparison of α-synuclein in PC12 cells. Scamp5 knockdown in PC12 cells resulted in α-synuclein accumulation. C Measurement of α-synuclein oligomers in PC12 cell by ELISA assay. D, E PC12 cells were transfected with the indicated siRNAs for 72 h, followed by flow cytometry analysis after staining with Propidium iodide and annexin V FITC. D The dot plot shows apoptosis and necrotic in control and treated cells. CON: control, UL: death cells, UR: late apoptosis cells, LR: early apoptosis cells, LL: normal cells. The red line indicates the position of the FITC line on the x-axis. E Quantitative measurement of the apoptosis cell. Data represent average population of representative experiments. F–I PC12 cells were transfected with indicated siRNAs for 72 h and protein levels were analyzed by Western blot. F, H Lysates were probed with indicated antibodies. The primary antibody was anti-Mcl-1, anti-Bcl2, anti-Bad, anti-caspase3, and anti-cleaved-caspase3. G, I Quantitative comparison of apoptosis-related factors in PC12 cells. Tubulin Beta 5 Class I (Tubulin5) served as a loading control. Data in panel B, E, G and I are presented as mean ± SEM. n ≥ 3 independent experiments. Significance levels are denoted as follows: ***P < 0.001