Figure 3

Exposure to 7-KC alters membrane order in CD4⁺T cells. Lymph node cells were exposed to di-4 ANEPPDHQ dye and stained with anti-Thy-1 AlexaFluor 647. T cells treated with 70 μM (C), 35 μM (D), 17.5 μM (E), mβCD-vehicle control (F) or left untreated (B) were gated (A) for Thy-1⁺ (FL4) and assessed for emission at 570 nm (FL2 channel) and 630 nm (FL3 channel) after appropriate compensation. T cells with high (lower most gate in B-F panels), intermediate (middle gate in B-F panels) and low (higher most gate in B-F panels) membrane order were enumerated and graphic representation of percent cells from four independent experiments with high, intermediate and low membrane order is shown in (G). Statistical significance between untreated, 7-KC treated groups was computed by two way ANOVA using JMP program. Different lower case alphabet designations (a – g) above each treatment group (none/untreated, mβCD, 17.5 μM, 35 μM, 70 μM 7KC) indicates statistically significant difference (p <0.0001) within the group of cells with high or intermediate or low membrane order. Similar lower case alphabet designation indicates lack of statistical significance (p > 0.8371).