annexin v apc pi dual staining apoptosis assay kit  (Abcam)

Journal: International Journal of Molecular Sciences

Article Title: Four and a Half LIM Domains 2 (FHL2) Contribute to the Epithelial Ovarian Cancer Carcinogenesis

doi: 10.3390/ijms21207751

Figure Lengend Snippet: Knockdown of FHL2 inhibits EOC cell growth and induced apoptosis. ( A ) Western blot results showing FHL2 expression levels in Hose 969, SKOV-3, IGROV-1, CAOV-3, COV-362, COV-644 and A2780 cell lines. Beta actin was used as a loading control. ( B ) FHL2 protein levels in SKOV–3 cells were transfected with 2 different FHL2 siRNAs (siFHL2-1, siFHL2-2). SKOV-3 cells transfected with scramble siRNA was used as control. Western blot results showed the FHL2 expression level in SKOV-3 control cells and FHL2 siRNAs knockdown cells. Beta actin was used as a loading control. ( C ) Morphological change of SKOV-3 cells transfected with scramble siRNA (Ctrl) or FHL2 siRNA (siFHL2). Scale bar: 100 μm. ( D ) SKOV-3 control cells and FHL2 knockdown cells (siFHL2) were stain with Annexin V-FITC/PI dual staining kit, and cell apoptosis was detected by the flow cytometry. Q1, Q2, Q3 and Q4 indicate proportion of dead cells, early apoptotic, live, and late apoptotic cells, respectively. Experiments were repeated at least 3 times, and representative images were shown. ( E ) MTT method results showed the cell viability changes in SKOV-3 control cells (Ctrl) and FHL2 knockdown cells transfected with 2 different siRNAs (siFHL2-1, siFHL2-2). Each bar represents mean ± S.E.M. (n = 3). ( F ) Cell number were quantified to show the FHL2 knockdown effect on SKOV-3 control cells (Ctrl) and FHL2 knockdown cells transfected with 2 different siRNAs (siFHL2-1, siFHL2-2). ( G ) Cell cycle distribution in SKOV–3 control (Ctrl) and FHL2 knockdown cells (siFHL2). Control and treated cells were labeled with propidium iodide and flow cytometry was used to determine the cell-cycle distribution. Each bar represents mean ± S.E.M. (n = 3). ** indicated p < 0.01 compared with control (Ctrl), *** indicated p < 0.001 compared with control (Ctrl).

Article Snippet: Apoptosis was analyzed by cell surface presence of Annexin V using the Annexin V-FITC/PI or Annexin V-APC/PI Dual Staining Apoptosis Assay Kit as described by the manufacturer (BioVision, Inc., Milpitas, CA, USA).

Techniques: Western Blot, Expressing, Transfection, Staining, Flow Cytometry, Labeling

Journal: International Journal of Molecular Sciences

Article Title: Four and a Half LIM Domains 2 (FHL2) Contribute to the Epithelial Ovarian Cancer Carcinogenesis

doi: 10.3390/ijms21207751

Figure Lengend Snippet: Ectopic expression of FHL2 promote EOC cell growth and induced cell morphological change. ( A ) IGROV-1 cells were transfected with letenvirus-based control vectors (Ctrl) or FHL2 overexpression vectors (FHL2 O/E) Beta actin was used as a loading control. ( B ) Representative images showed the morphological change of IGROV-1 control cells (Ctrl) and FHL2 overexpressing cells (FHL2 O/E). Scale bar: 100 μm. ( C ) Growth curve of IGROV-1 control cells (Ctrl) and FHL2 overexpressing cells (FHL2). Each point represents mean ± S.E.M. *** indicate p < 0.001. ( D ) Quantification results showed the apoptotic, live and dead cell proportion in IGROV-1 control (Ctrl) and FHL2 overexpression cells (FHL2 O/E). ( E ) IGROV-1 control (Ctrl) and FHL2 overexpression cells (FHL2 O/E) cells were cultured in serum reduced (2%) culture medium. Cells then stained with Annexin V-APC/PI dual staining kit for flowcytometry analysis.

Article Snippet: Apoptosis was analyzed by cell surface presence of Annexin V using the Annexin V-FITC/PI or Annexin V-APC/PI Dual Staining Apoptosis Assay Kit as described by the manufacturer (BioVision, Inc., Milpitas, CA, USA).

Techniques: Expressing, Transfection, Over Expression, Cell Culture, Staining

Journal: International Journal of Molecular Sciences

Article Title: Four and a Half LIM Domains 2 (FHL2) Contribute to the Epithelial Ovarian Cancer Carcinogenesis

doi: 10.3390/ijms21207751

Figure Lengend Snippet: FHL2 regulate AKT expression in EOC cells. ( A ) Western blot results showed FHL2, AKT and apoptosis related protein levels in SKOV-3 control and FHL2 knockdown cells (siFHL2). Beta tubulin was used as loading control. ( B ) Western blot results showed FHL2 and AKT expression level in IGROV-1 control and FHL2 overexpression cells (FHL2). Beta actin was used as loading control.

Article Snippet: Apoptosis was analyzed by cell surface presence of Annexin V using the Annexin V-FITC/PI or Annexin V-APC/PI Dual Staining Apoptosis Assay Kit as described by the manufacturer (BioVision, Inc., Milpitas, CA, USA).

Techniques: Expressing, Western Blot, Over Expression