Journal: Frontiers in Immunology
Article Title: Altered Metabolic Phenotype of Immune Cells in a Spontaneous Autoimmune Uveitis Model
doi: 10.3389/fimmu.2021.601619
Figure Lengend Snippet: ATP5β in CD4 + T cells of healthy and ERU cases. (A) Western blot with CD4 + T cells of control and ERU horses (n=5 each) stained with anti ATP5β antibody, anti GAPDH antibody and anti β-actin antibody for normalization. A molecular weight marker is shown on the left, followed by a blank lane and a lane only incubated with the secondary antibody. (B) Statistical analysis of the volume of ATP5β lanes of control and ERU cases after normalization to the respective β-actin lane showed no significant difference (ns p > 0.05). Data are represented as mean ± SEM. The ERU bar (blue) was set accordingly to the control bar (grey), which was set as 1. (C) Representative immunocytochemically stained CD4 + T cell of a control (upper picture) and an ERU case (lower picture). Mitochondria were stained with MitoTracker Deep Red (red). Cell nuclei were counterstained with DAPI (blue). x1000 magnification.
Article Snippet: Purity of CD4 + fraction was tested via flow cytometry, by staining with 30 μl mouse anti horse CD4 FITC labelled antibody (clone MCA1078F, Biorad, Feldkirchen, Germany, 1:10) and 30 μl mouse anti horse CD8 FITC labelled antibody (clone MCA1080F, Biorad, Feldkirchen, Germany, 1:10) ( , ≥ 96% purity of the CD4 + fraction).
Techniques: Western Blot, Control, Staining, Molecular Weight, Marker, Incubation