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. Author manuscript; available in PMC: 2016 Nov 4.
Published in final edited form as: J Immunol. 2014 Jun 4;193(1):412–421. doi: 10.4049/jimmunol.1400055

Figure 4. TCR activation increases FasL expression and cell death in control and ethanol treated CD4+ T lymphocytes.

Figure 4

Freshly isolated CD4+ T cells from healthy individuals (n=3) were untreated (UT) or exposed to 25mM ethanol (E) for 24h and then subsequently stimulated with 1μg/ml anti CD3/CD28 antibody (S and E+S). (A)Total cellular lysates (24 h post stimulation) were analyzed for FasL protein expression by Western blot analysis with β-actin as loading control. A representative of 3 separate experiments done on CD4+ T cells obtained from different individuals is shown. Densitometry analysis was done and the ratios were noted at bottom. (B) DNA fragmentation analysis was performed following treatment with human neutralizing Anti-Fas Ligand (10μg/ml, anti-Fas) or isotype IgG control (10μg/ml, IgG) antibodies in control (Clear bar) or ethanol (Gray bar) treated cells. P values: * = p<0.01 when comparing control baseline to control stimulation, ** = p<0.001 when comparing control stimulation to Ethanol +Stimulation, ‡ = p<0.001 when comparing Ethanol +Stimulation with Ethanol + anti-FasL+S, ϯ = p<0.001 when comparing for anti-FasL+S with IgG+S, ϯϯ = p<0.01 when comparing E+anti-FasL+S with E+IgG+S. Data represents mean ± SE (n = 3).