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. 2013 Jul 2;27(9):1455–1467. doi: 10.1210/me.2013-1075

Figure 2.

Figure 2.

Poly(I:C)-induced expression of cytokines. A, Expression of TLR3, MDA5, and RIG-I. Leydig and germ cells were isolated from 5-week-old WT and TLR3-deficient (TLR3−/−) mice. The absence of TLR3 mRNA in TLR3−/− cells was confirmed by real-time qRT-PCR (left panel). The proteins of TLR3, MDA5, and RIG-I in WT and TLR3−/− cells were determined by Western blot (right panel). B and C, Poly(I:C)-induced up-regulation of cytokines. Total RNA was extracted from TLR3−/− Leydig and germ cells after transfection with poly(I:C) for the specified duration. The mRNA levels of the indicated cytokines were analyzed using real-time qRT-PCR. D, Cytokine secretion in culture media. The cells were seeded in 6-well plates at a density of 5 × 105 Leydig cells or 1 × 106 germ cells per well and transfected with poly(I:C). After 24 hours, the media were measured for cytokine concentration using ELISA. Data are the mean ± SEM of the 3 experiments. *, P < .05; **, P < .01. ND, not detectable.