Evidence-Based Complementary and Alternative Medicine / 2013 / Article / Fig 3

Research Article

Reishi Protein LZ-8 Induces FOXP3+ Treg Expansion via a CD45-Dependent Signaling Pathway and Alleviates Acute Intestinal Inflammation in Mice

Figure 3

LZ-8 promotes Treg expansion via a CD45-mediated signal pathway. (a) Human T-cell Jurkat (WT), J.RT3-T3.5 (TCR−/−), and J45.01 (CD45−/−) cells were cultured without stimulation as the control (CTR), with stimulation by rLZ-8 (1, 2 μg/mL), or with stimulation by anti-CD3/28 antibodies (CD3/CD28; 1,  2 μg/mL) for 72 h. The cells were then harvested for flow cytometry analysis of CD25 and FOXP3 expression. (b) J.RT3-T3.5 cells were stimulated with or without rLZ-8 (2 μg/mL) or with or without anti-CD3/28 antibodies (CD3/28; 2 μg/mL) for the indicated time periods. The total cell lysates were collected for protein phosphorylation analysis. (c) Primary human CD4+ T cells were pretreated with or without CD45 phosphatase inhibitor (CD45i; 1, 3 μM) for one hour prior to rLZ-8 stimulation for 72 h. The cells were then harvested for flow cytometry analysis of CD25 and FOXP3 expression.
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