Fig. 6

Effect of palmitoleic acid (PA) and oleic acid (OA) at non-toxic concentration on T regulatory cell differentiation. Cells were incubated with 25 and 50 μM of PA or OA in the presence or absence of ConA for 24 h. After, cells were stained with anti-CD4 (FITC) and anti-CD25 (APC). The negative control cells were incubated with non-reactive labeled antibody IgG. After this period, the cells were fixed and incubated in buffer containing a permeabilizing agent followed by incubation with anti-Foxp3 (PE). Twenty thousand events in CD4+ cells per sample were acquired in histograms on the flow cytometer BD-Accuri. At first, a determination of CD4+ cells was performed and from these, the percentage of CD25 + Foxp3 + cells was determined. Representative dot plots of APC fluorescence (x axis) and PE fluorescence (y axis) used as indicators of CD25 and Foxp3 are shown in: a Control cells; b ConA stimulated control cells; c 50 μM PA treated cells; d 50 μM PA + ConA treated cells; e 50 μM OA treated cells; f 50 μM OA + ConA treated cells The values are presented as mean ± S.E.M. of three determinations from six experiments. *P < 0.01 versus control