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. 2011 Mar;132(3):361–375. doi: 10.1111/j.1365-2567.2010.03369.x

Figure 3.

Figure 3

Thy-1 cross-linking enhances α-galactosylceramide (αGC)-mediated cytokine production by mouse CD4−CD8− invariant natural killer (iNKT) cells. DN32.D3 cells were incubated with αGC alone (100 ng/ml), with a low (5 μg/ml) or high (40 μg/ml) dose of G7, or with a combination of αGC and G7. After 24 hr, culture supernatants were harvested and interleukin-2 (IL-2), interferon-γ (IFN-γ) and IL-4 levels were measured by ELISA (a). After 24 hr, cells were harvested and total RNA was extracted. RNA was reverse transcribed into cDNA, which was then amplified by quantitative PCR using primers specific for IL-2, IFN-γ and IL-4. Cytokine mRNA levels were normalized to glyceraldehyde 3-phosphate dehydrogenase and the normalized values were graphed as expression ratios. Expression ratios were relative to untreated cells, which were assigned a value of 1 (b). Statistical significance is denoted by asterisks, where ** and *** represent P < 0·01 and P < 0·001, respectively.