Bio Background |
NIP3 is a member of the BCL2/adenovirus E1B 19 kd-interacting protein (BNIP) family. It interacts with the E1B 19 kDa protein which is responsible for the protection of virally-induced cell death, as well as E1B 19 kDa-like sequences of BCL2, also an apoptotic protector. NIP3 contains a BH3 domain and a transmembrane domain, which have been associated with pro-apoptotic function. The dimeric mitochondrial protein is known to induce apoptosis, even in the presence of BCL2. |
Bio References |
References for protein:1.Kothari, S., et al., Oncogene 22(30):4734-4744 (2003). 2.Lee, S.M., et al., Life Sci. 71(19):2267-2277 (2002). 3.Ray, R., et al., J. Biol. Chem. 275(2):1439-1448 (2000). 4.Chen, G., et al., J. Biol. Chem. 274(1):7-10 (1999). 5.Yasuda, M., et al., J. Biol. Chem. 273(20):12415-12421 (1998).References for HepG2 cell line:1. Knowles BB, et al. (1980). Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science 209: 497-499.[ PubMed: 6248960].2. Darlington GJ, et al. (1987). Growth and hepatospecific gene expression of human hepatoma cells in a defined medium. In Vitro Cell. Dev. Biol. 23: 349-354.[PubMed: 3034851].3. Ihrke, G; Neufeld, EB; Meads, T; Shanks, MR; Cassio, D; Laurent, M; Schroer, TA; Pagano, RE et al. (1993). "WIF-B cells: an in vitro model for studies of hepatocyte polarity". Journal of Cell Biology 123 (6): 1761–1775. [PubMed:7506266]. 4. Mersch-Sundermann, V.; Knasmüller, S.; Wu, X. J.; Darroudi, F.; Kassie, F. (2004). "Use of a human-derived liver cell line for the detection of cytoprotective, antigenotoxic and cogenotoxic agents". Toxicology 198 (1–3): 329–340. [PubMed:15138059]. |
Legend image 1 |
Fluorescent confocal image of HepG2 cells stained with BNIP3 (BH3 Domain Specific) antibody. HepG2 cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with AP1321a BNIP3 (BH3 Domain Specific) primary antibody (1:500, 2 h at room temperature). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min). BNIP3 immunoreactivity is localized to the cytoplasm of HepG2 cells. |
Legend image 2 |
Freshly isolated mouse hepatocytes plated on coverslips (2 x105 cells/22-mm glass coverslip) were cultured under normoxic conditions for 6 hr. The cells were then fixed in 2% paraformaldehyde in PBS for 1 hr, and processed for confocal immunofluorescence (red: F-actin, blue: ATP-synthase, green: BNIP3). Fluorescence labeling of BNIP3 accomplished with anti-BNIP3 antibody Cat # AP1321a. Data courtesy of Ruben Zamora, University of Pittsburgh. |