Journal: Heliyon

Article Title: Insulin attenuates apoptosis in neuronal cells by an integrin-linked kinase-dependent mechanism

doi: 10.1016/j.heliyon.2019.e02294

Figure Lengend Snippet: Insulin increases IAP expression in an ECM-dependent manner in Cerebellar Granule Neurons: Inhibition by the ILK Inhibitor KP-392. Representative Western blots of lysates from cerebellar cultures grown on poly-D-lysine or laminin and exposed to serum and depolarization or serum and depolarization withdrawal alone or in the presence of insulin (10 μg/ml) with or without KP-392 (100 μM) for 24 h (n = 2–3). Immunoblots were probed with an antibody against c-IAP2. Bar graph represents densitometric analysis of 3 independent trials of cells grown on laminin, averaged and normalized to cultures subject to serum and depolarization withdrawal. Data are mean ± SEM. *P < 0.05, different from insulin treated group as determined by a Student's t-test. Full, non-adjusted images are not available as supplementary material for Fig. 5 .

Article Snippet: Lysates from primary cultures were prepared in Tris-HCl buffer, pH 7.6 containing 1% NP-40, 150 mM NaCl, 1mM EDTA, 3.8 μg/ml aprotinin, 1 μg/ml leupeptin, 1 mM PMSF, 2 mM NaF, and 1 mM Na 3 VO 4 as previously described . c-IAP2 was detected using a rabbit anti-c-IAP2 antibody (a kind gift from Dr. Robert Korneluk, Aegera Therapeutics Inc., QC, CA), ILK was detected using a monoclonal anti-ILK antibody (BD Transduction Laboratories, San Jose, CA), caspase was detected using a monoclonal antibody caspase-3 p11 (Santa Cruz, ON, CA) and GAPDH was detected using a rabbit anti-GAPDH (Santa Cruz, ON, CA).

Techniques: Expressing, Inhibition, Western Blot