Journal: Cells
Article Title: Cys340Ser Mutation Abolishing S-Nitrosylation Drives GRK2 Mitochondrial Localization and Dysfunction
doi: 10.3390/cells15050458
Figure Lengend Snippet: Group comparison of Mfn1 ( A ), Mfn2 ( B ) and Opa1 ( C ) expressions. Statistical significance was determined by ANOVA followed by Bonferroni’s post hoc test. Data are shown as mean ± SEM; ns, p = 0.4591 for Mfn1; p = 0.1796 for Mfn2; p = 0.0552 for Opa1 ( n = 7–9 per group). Representative immunoblots demonstrating Mfn1, Mfn2 and Opa1 protein levels in total cell lysates of AC16 cells infected with Ad.GFP or Ad.GRK2-C340S under normoxic or hypoxia/reoxygenation (H/R) conditions. GAPDH served as a total protein marker. Densitometric analysis was performed using Li-Cor Image Studio software and normalized to the respective loading controls. All samples included in each comparative analysis were run on the same gel. Cropped blots are presented for clarity; uncropped blots with molecular weight markers are provided in .
Article Snippet: The primary antibodies used were as follows: GRK2 (13990-1-AP; ProteinTech, Rosemont, IL, USA), GRK2 (05-465; Sigma Aldrich, St. Louis, MO, USA), Drp1 (8570; Cell Signaling Technology, Danvers, MA, USA), Fis1 (32525; Cell Signaling Technology, Danvers, MA, USA), Mfn1 (14739; Cell Signaling Technology, Danvers, MA, USA), Mfn2 (11925; Cell Signaling Technology, Danvers, MA, USA), Opa1 (80471; Cell Signaling Technology), Pink (6946; Cell Signaling Technology, Danvers, MA, USA), Parkin (4211; Cell Signaling Technology, Danvers, MA, USA), LC3 (3868; Cell Signaling Technology, Danvers, MA, USA), Tom20 (42406; Cell Signaling Technology, Danvers, MA, USA), VDAC1 (sc-390996; Santa Cruz Biotechnology, Dallas, TX, USA), and GAPDH (sc-32233; Santa Cruz Santa Cruz Biotechnology, Dallas, TX, USA).
Techniques: Comparison, Western Blot, Infection, Marker, Software, Molecular Weight
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