Figure 5
Assessment of viability of CD4+T cells treated with 7-Keto cholesterol. A. Lymph node cells were cultured with 35 μM, 17.5 μM 7-KC, mβCB vehicle control either in the presence of c-Ova323–339 peptide (Panel B) or anti-CD3ε monoclonal antibody (C). Viable cells were enumerated by staining with CFSE (X-axis) and Propidium Iodide (Y-axis). Live cells in these cell cultures were quantified by enumerating CFSEhigh PInegative cells. These data graphically represented in panels B & C are obtained from three independent experiments. A representative flow analysis of one of the three experiments computed for panel C is shown in panel A. Statistical significance between treated and untreated groups was computed by one way ANOVA using the JMP program. Similar lower case alphabet designations (a) above each treatment group (control, 35 μM 7KC, 17.5 μM 7KC, mβCD) indicates lack of statistically significant differences (p = 0.21 for cOva peptide responses and p = 0.06 for anti-CD3ε responses). To examine statistical significance within a specific antigen receptor response (cOva323–339 or anti-CD3ε), each treatment group (e.g., 7KC) is compared with other (all possible combinations).