Journal: Journal of Lipid Research

Article Title: Lactosylceramide contributes to mitochondrial dysfunction in diabetes

doi: 10.1194/jlr.m060061

Figure Lengend Snippet: Fig. 1. Heart mitochondria display minimal contamination with other cellular membrane compartments. Western blots for Na + ,K + - ATPase (plasma membrane marker), LAMP-2 (lysosomal marker), calnexin (ER marker), syntaxin 6 (Golgi marker), and VDAC (mi- tochondrial marker) were performed by loading equal amounts of 30 g of homogenate (H), or mitochondrial (SSM or IFM) protein per lane, except VDAC, which was assessed using protein loading of 15 g per lane. The blot is representative of three independent experiments .

Article Snippet: Antibodies against mitochondrial marker, voltage dependent anion channel (VDAC) (D73D12), and syntaxin 6 (Golgi marker) were supplied by Cell Signaling Technology (Danvers, MA).

Techniques: Membrane, Western Blot, Clinical Proteomics, Marker

Journal: Journal of Lipid Research

Article Title: Lactosylceramide contributes to mitochondrial dysfunction in diabetes

doi: 10.1194/jlr.m060061

Figure Lengend Snippet: Fig. 4. Effect of diabetes on the abundance of ceramide metabolizing enzymes in heart lysates (A, B) and isolated mitochondria (C, D). Western blots were performed by loading equal amounts of 30 g of homogenate (A), or mitochondrial (SSM or IFM) protein per lane, except VDAC, which was assessed using protein loading of 15 g per lane. A: Representative Western blot of heart lysates. B: Quantifi cation of protein expression in heart lysates. Levels of respective proteins were normalized against -actin and their expression in STZ animals relative to control was determined. Data are mean ± SE of three homogenate preparations; eight to ten mice were used for each prepara- tion of homogenate. * P < 0.05. C: Representative Western blot of heart mitochondria. D: Quantifi cation of protein expression in heart mitochondria. Levels of respective proteins were normalized against VDAC and their expression relative to IFM control was determined. Data are mean ± SE of three preparations; eight to ten mice were used for each preparation of mitochondria. * P < 0.05.

Article Snippet: Antibodies against mitochondrial marker, voltage dependent anion channel (VDAC) (D73D12), and syntaxin 6 (Golgi marker) were supplied by Cell Signaling Technology (Danvers, MA).

Techniques: Isolation, Western Blot, Expressing, Control