Journal: Molecular Medicine Reports

Article Title: Effects of nuclear respiratory factor-1 on apoptosis and mitochondrial dysfunction induced by cobalt chloride in H9C2 cells

doi: 10.3892/mmr.2019.9839

Figure Lengend Snippet: Effects of CoCl 2 on apoptosis. (A) Changes in nuclear morphology, representative of apoptosis, in cells induced by CoCl 2 (400 µM) for 6 h were observed by Hoechst 33342 staining in pLenti-H9C2 and NRF1-H9C2 cells. Cells with intense blue staining in nuclei were counted. Magnification, ×400. (B) Flow cytometry was used to detect apoptosis following staining with Annexin V-APC and 7-ADD. (C) mRNA expression of Bcl-2 and Bax was analyzed by reverse transcription-quantitative polymerase chain reaction. (D) Protein expression levels of NRF-1, Bcl-2, Bax and PARP were analyzed by western blot. **P<0.01 vs. NC; # P<0.05, ## P<0.01 vs. pLenti-H9C2 cells. NC, negative control (CoCl 2 untreated cells); CoCl 2 , cobalt chloride; NRF, nuclear respiratory factor; 7-AAD, 7-aminoactinomycin D; APC, allophycocyanin; Bcl-2, B-cell lymphoma-2; Bax, Bcl-associated X protein; PARP, poly(ADP-ribose) polymerase 1.

Article Snippet: Cells were seeded in 24-well plates (2×10 5 per well) and cultured with medium containing 400 µM CoCl 2 for 6 h. Apoptotic cells were measured by flow cytometry using Annexin V-APC and 7-AAD Apoptosis Detection reagent (BD Biosciences).

Techniques: Staining, Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Negative Control