Journal:

Article Title: Essential Role of the Redox Sensitive Kinase p66 shc in Determining Energetic and Oxidative Status and Cell Fate in Neuronal Preconditioning

doi: 10.1523/JNEUROSCI.6366-09.2010

Figure Lengend Snippet: Cultures were fixed 24h after the initiation of preconditioning and stained with the nuclear dye DAPi (blue), the neuronal cytoskeletal marker MAP2 (green) and LC3 (red), a hallmark of autophagic activation. A) Control cultures had well defined processes and light dispersed LC3 activation. B) Autophagosome punctae containing LC3 increased in response to preconditioning (bottom left panel in red). C) Exposure to the Shci PP2 (50nM) exacerbated this effect. D) Cell counts by two independent investigators of > 400 neurons from three independent experiments revealed a 20% increase in intensely labeled LC3II positive cells in preconditioned cells and a further increase of 15% when Shci was present.

Article Snippet: Primary antibodies used in these studies were phospho-ERK (4377, Cell Signaling, Danvers, MA), HSP70 and HSC70 (SPA-811 and SPA-816, Assay Designs, Ann Arbor, Michigan), phospho p66 Shc (Calbiochem, San Diego, CA), phospho Raf S338 (Santa Cruz, Santa Cruz, CA), LC3 (PD014, MBL International Corp, Woburn, MA), and MAP2 (M4403, Sigma, St. Louis, MO).

Techniques: Staining, Marker, Activation Assay, Control, Labeling