Fig 1: Overexpression of Septin4 promotes apoptosis in hFOB 1.19 cells treated with a high melatonin concentration. (A) hFOB cells were transfected with NC plasmid or Flag-Septin4 plasmid for 48 h and treated with 0, 2, 4 and 6 mmol/l of melatonin for 48 h. Cell viability was assessed via the MTT assay. (B) hFOB cells were transfected with NC plasmid or Flag-Septin4 plasmid for 48 h and treated with 0 or 4 mmol/l of melatonin for 48 h. Flow cytometry was performed to analysis cell apoptosis (Q4 region). *P<0.05, **P<0.01, ***P<0.001, vs. NC group. GRP, glucose-regulated protein; NC, negative control; Con, control; PI, propidium iodide.
Fig 2: Septin4 participated in DOX-induced apoptosis of HCT116 cells. A. HCT116 cells were treated with DOX at 0, 0.005, 0.05, 0.2, 0.4, and 2 μmol/L for 48 h, to detect the expression of cleaved-caspase3 and cleaved-PARP1. B. Quantification of protein expression in A were shown as the means±S.D., ***P<0.001 C. The expression of PCNA and Septin4 was detected in the presence of DOX at 0, 0.005, 0.05, 0.2, 0.4, and 2 μmol/L for 48 h. D. Quantification of protein expression in C were shown as the means±S.D., **P<0.01, ***P<0.001. E. The generation of ROS was detected by DHR staining under the treatment of 0, 0.005, 0.05, 0.2, 0.4, and 2 μmol/L DOX for 48 h. Scale bars, 100 μm. F. Quantitative analyses of the fluorescence intensity in E, data are shown as means±S.D., ***P<0.001. G. Statistical results of CCK8 analysis of HCT116 treated with DOX at 0, 0.005, 0.05, 0.2, 0.4, and 2 μmol/L for 48 h, data are shown as means±S.D., ***P<0.001.
Fig 3: Effect of Septin4 knockdown and overexpression on PPARγ, LXRα and ABCA1/G1 expression in THP-1 macrophage-derived foam cells. (A) ABCA1, ABCG1, (B) PPARγ and LXRα protein expression levels in THP-1 macrophage-derived foam cells following Septin 4 knockdown or overexpression were analyzed using western blotting. **P<0.01 and ***P<0.001. PPARγ, proliferator activated receptor γ; LXRα, liver X receptor α; ABCA1, ATP binding cassette subfamily A member 1; ABCG1, ATP binding cassette subfamily G member 1; NC, negative control; sh, short hairpin; ov, overexpression; ev, empty vector.
Fig 4: Overexpression of Septin4 increases ERS- and apoptosis pathway-related gene expression levels in hFOB 1.19 cells treated with a high melatonin concentration. hFOB 1.19 cells were transfected with Flag-Septin4 plasmid or NC plasmid for 48 h and treated with 0 or 4 mmol/l of melatonin for 48 h and western blotting was performed to detect the expression levels of Flag, Septin4, GRP78, GRP94 and cleaved caspase-3. *P<0.05, **P<0.01, ***P<0.001, vs. NC group. GRP, glucose-regulated protein; NC, negative control; Con, control.
Fig 5: Expression of Septin4 is increased in hFOB 1.19 cells treated with a high concentration of melatonin. (A) hFOB 1.19 cells were treated with 0, 2, 4 and 6 mmol/l of melatonin for 48 h. Cell viability was assessed via the MTT assay. (B) Flow cytometry was performed to analyze apoptosis in hFOB 1.19 cells, which were treated with 0, 2, 4 and 6 mmol/l of melatonin. (C) Western blotting was performed to detect the expression levels of GRP78, GRP94, cleaved caspase-3 and Septin4 after treatment with 0, 2, 4 and 6 mmol/l of melatonin for 48 h. *P<0.05, **P<0.01, ***P<0.001 vs. 0 mmol/l group. GRP, glucose regulated protein; PI propidium iodide.
Supplier Page from Abcam for Anti-SEPT4 antibody