primary antibodies (mfn2, grp78, perk, p-perk, grp75, micu1, ip3r1, mcu, vdac1, and β-actin)  (Cell Signaling Technology Inc)

Journal: Biomolecules

Article Title: Resveratrol Treatment in Human Parkin -Mutant Fibroblasts Modulates cAMP and Calcium Homeostasis Regulating the Expression of Mitochondria-Associated Membranes Resident Proteins

doi: 10.3390/biom11101511

Figure Lengend Snippet: Effect of resveratrol on protein levels of GRP75, Miro2, and Mfn2 in control and parkin -mutant fibroblasts. ( a ) Representative image of Western blot of GRP75, Miro2, and Mfn2 performed on whole cell lysates from control and parkin -mutant fibroblasts exposed to either vehicle (DMSO) or 25 μM resveratrol for 24 h (MW, molecular weight). The graphs (panel ( b ), GRP75; panel ( c ), Miro2; panel ( d ), Mfn2) display the statistical densitometric analysis of band intensity of proteins normalized to the corresponding actin level, used as loading control. Data means ± SEM from 3 independent experiments under each condition are expressed as percentage of vehicle-treated control cells. The significance was determined by one-way ANOVA with Tukey post hoc; ***, p ˂ 0.0005; ****, p ˂ 0.0001. For more details, see Materials and Methods.

Article Snippet: Total cell proteins (20 μg) from control and parkin -mutant fibroblasts exposed to either vehicle (0.02% DMSO) or 25 μM resveratrol (RSV) for 24 h were separated on a 8% Tris-Glycine SDS–PAGE, transferred to nitrocellulose membranes with 0.2 µm pore size (Bio-Rad, Hercules, CA, USA), and immunoblotted with specified primary antibodies against GRP75 (1:200; Santa Cruz Bio Technology, Dallas, TX, USA, Catalog number: sc-13967), Miro2 (1:1000; Cell Signaling Technology, Danvers, MA, USA, Catalog number: #14016), and Mfn2 (1:200; Millipore, Burlington, MA, USA, Catalog number: #ABC42).

Techniques: Mutagenesis, Western Blot, Molecular Weight