Figure 6.

Gβγ inhibition decreases mRNAs encoding STAT4 and some STAT4-regulated genes in CD4⁺ TH2 memory cells. (A) Gallein, but not fluorescein, and Gβ₁ siRNA significantly decreased levels of mRNA encoding STAT4 (A), LAG-3 (B), and NKG-7 (C). (D) Gallein decreased MAP3K8 mRNA levels significantly relative to fluorescein-treated cells, but not to control cells. Gβ₁ siRNA did not decrease MAP3K8 mRNA levels. (A-D) First set of graphs: box plots (top) and difference plots (bottom) show data from memory CD4⁺ T cells isolated from the peripheral blood of 20 healthy donors, stimulated with plate-bound anti-CD3 and soluble anti-CD28, and grown in conditions promoting TH2 differentiation in the absence or presence of gallein or fluorescein for three days. Second set of graphs: box plots (top) and difference plots (bottom) show data from memory CD4⁺ T cells isolated from the blood of 30 healthy donors and stimulated for three days with plate-bound anti-CD3 and soluble anti-CD28 in conditions promoting TH2 differentiation in the presence of Gβ₁ siRNA or NT siRNA. mRNA levels were determined by qPCR. ^*, p < 0.05; ^**, p < 0.01; ^***, p < 0.001; ^****, p < 0.0001.