Journal: Molecular Cancer

Article Title: Matrine induces caspase-independent program cell death in hepatocellular carcinoma through bid-mediated nuclear translocation of apoptosis inducing factor

doi: 10.1186/1476-4598-13-59

Figure Lengend Snippet: AIF translocation is required for matrine - induced ciPCD. (A-B) HepG2 cells were transfected with AIF siRNA (40 or 60 nM) or non-targeted siRNA for 24 hrs, and then treated with matrine at 1.5 mg/ml for 24 hrs. (A) AIF expression levels were detected by western blot analysis using an anti-AIF antibody. Actin was used as a protein loading control. (B) Cells were stained with Annexin V/PI to assess cell viability by flow cytometry. The basal level of cell death was normalized to 1.* p < 0.05, ** p < 0.01 vs . non-target siRNA + Matrine. (C) Immunostaining was performed to localize AIF with an anti-AIF antibody and a FITC-conjugated secondary antibody (green). The nuclei were detected using DAPI staining (blue). (D) Cytoplasmic, nuclear, and mitochondrial fractions were prepared for western blot analysis using anti-AIF, anti-β-tubulin (cytosolic marker), anti-Hsp60 (mitochondrial marker) and anti-Lamin B (nuclear marker) antibodies.

Article Snippet: Bid siRNA, AIF siRNA and nonfunctional control siRNA were purchased at Santa Cruz Biotechnology.

Techniques: Translocation Assay, Transfection, Expressing, Western Blot, Control, Staining, Flow Cytometry, Immunostaining, Marker

Journal: Molecular Cancer

Article Title: Matrine induces caspase-independent program cell death in hepatocellular carcinoma through bid-mediated nuclear translocation of apoptosis inducing factor

doi: 10.1186/1476-4598-13-59

Figure Lengend Snippet: Bid mediated the cell death induced by matrine , as an upstream regulator of AIF. HepG2 cells were transfected with Bid siRNA (40 or 60 nM) or non-targeted siRNA for 24 hrs (A , C , E) or pretreated Bid inhibitor BI-6C9 (10 μM) for 1 hr (B, D, F) , then treated with matrine at 1.5 mg/ml for 24 hrs. (A , B) Immunostaining was performed to localize AIF with an anti-AIF antibody and a FITC-conjugated secondary antibody (green). The nuclei were detected using DAPI staining (blue). (C , D) Cytoplasmic, nuclear, and mitochondrial fractions were prepared for western blot analysis to localize AIF with an anti-AIF antibody using anti-AIF, anti-β-tubulin (cytosolic marker), anti-Hsp60 (mitochondrial marker) and anti-Lamin B (nuclear marker) antibodies. (E , F) Cells were stained with Annexin V/PI to assess cell viability by flow cytometry. The basal level of cell death was normalized to 1.* p < 0.05, ** p < 0.01 vs . non-target siRNA + Matrine (E) , Matrine (F) .

Article Snippet: Bid siRNA, AIF siRNA and nonfunctional control siRNA were purchased at Santa Cruz Biotechnology.

Techniques: Transfection, Immunostaining, Staining, Western Blot, Marker, Flow Cytometry