miR-155, a Modulator of FOXO3a Protein Expression, Is Underexpressed and Cannot Be Upregulated by Stimulation of HOZOT, a Line of Multifunctional Treg
Figure 2
Underexpression and non-responsiveness of miR-155 are unique features of HOZOT and correlated with FOXP3 expression.
A. Decreased expression of miR-155 in HOZOT was validated by real-time PCR using HOZOT cell lines (n = 5) in comparison with Tconv cell lines (n = 5) and Treg cell lines (n = 3). All RNA samples were prepared from the cell lines before re-stimulation. The ratio of miR-155/U6 small nuclear RNA (snRNA) in HOZOT-1 was defined as 1.0. B. miR-155 expression does not respond to stimulation in HOZOT. HOZOT (n = 5), Tconv (n = 3), and Treg (n = 3) cells were re-stimulated with CD3/CD28. miR-155 expression was monitored at zero, three and 24 hrs after re-stimulation. The ratio of miR-155/U6 snRNA in HOZOT was defined as 1.0. C. Expression of FOXP3 in HOZOT is not responsive to stimulation. Flow cytometric analysis of FOXP3 expression was assayed by intracellular staining 24 hrs after re-stimulation. Values in histograms indicate the percentage of FOXP3+ cells.