Effect of IL-36R ligands on mRNA expression of soluble and cell-associated inflammatory mediator by BMDCs
| . | Unstimulated: Ct . | IL-36Ra . | IL-36α . | IL-36β . | IL-36γ . | IL-1β . | LPS . | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Ct . | Fold . | Ct . | Fold . | Ct . | Fold . | Ct . | Fold . | Ct . | Fold . | Ct . | Fold . | ||
| IL-6 | 29.7 ± 1.8 | 29.9 ± 1.3 | 1.0 | 21.3 ± 0.8 | 218.3 | 21.6 ± 2 | 560 | 21 ± 0.8 | 311.5 | 27.8 ± 1 | 3.3 | 18.5 ± 0.4 | 1216.2 |
| IL-12p40 | 31.1 ± 0.9 | 31.5 ± 0.8 | 0.0 | 25 ± 1.4 | 139.6 | 23.6 ± 1.5 | 426.7 | 22.8 ± 2.1 | 388 | 30 ± 1.3 | 2.5 | 20 ± 1.2 | 1638.8 |
| COX-2 | 29.5 ± 0.2 | 29.8 ± 0.3 | 1 | 22.1 ± 0.5 | 146.5 | 22.6 ± 2.7 | 322 | 21.6 ± 0.6 | 238.7 | 28 ± 0.1 | 2.9 | 20.4 ± 0.4 | 384.5 |
| CXCL1 | 28.5 ± 0.6 | 28.5 ± 0.4 | 1.1 | 20.5 ± 0.5 | 208.9 | 19.9 ± 0.3 | 320.3 | 20.4 ± 0.5 | 239.2 | 25.6 ± 0.3 | 7 | 19.6 ± 0.6 | 297 |
| CCL1 | 33.3 ± 0.9 | 33.1 ± 0.8 | 1.4 | 27.6 ± 2.4 | 68.4 | 27 ± 0.8 | 270.2 | 26 ± 2.2 | 175.4 | 31.4 ± 1.5 | 4.2 | 26.1 ± 1.3 | 107.6 |
| IL-12p35 | 34.3 ± 1.2 | 34.2 ± 0.8 | 1.5 | 27.8 ± 0.9 | 86 | 28 ± 1.7 | 216.5 | 27.4 ± 0.9 | 129.5 | 32.7 ± 1.3 | 4.5 | 27 ± 0.6 | 116.5 |
| IL-1β | 25.1 ± 0.9 | 25.2 ± 0.6 | 1.1 | 17.7 ± 0.6 | 139 | 18.7 ± 2.3 | 211.3 | 17.5 ± 0.6 | 171.6 | 23 ± 0.75 | 5 | 16.9 ± 0.5 | 185 |
| IL-23p19 | 30 ± 1.1 | 30.3 ± 0.9 | 1 | 24 ± 1.2 | 53 | 22.2 ± 0.9 | 207.4 | 23.2 ± 1 | 97.2 | 28.5 ± 0.7 | 2.9 | 22.4 ± 1 | 134.8 |
| GM-CSF | 33 ± 0.7 | 32.6 ± 0.9 | 2 | 27.6 ± 0.8 | 40.6 | 27.8 ± 2 | 111.2 | 26.5 ± 0.7 | 88.2 | 29.5 ± 0.6 | 13.4 | 29 ± 0.5 | 11.8 |
| IL-10 | 34.5 ± 1 | 40 ± 0.0 | 0 | 31.2 ± 0.5 | 16 | 30.4 ± 2.8 | 91 | 29.7 ± 0.7 | 49 | 34.9 ± 0.3 | 2 | 29.5 ± 0.1 | 42 |
| CXCL10 | 28 ± 1.7 | 28 ± 1.5 | 1.5 | 25.8 ± 1.4 | 3.9 | 25.5 ± 4.3 | 47 | 25.5 ± 1.4 | 5 | 27.1 ± 1 | 2 | 18.25 ± 0.7 | 644.3 |
| NOS2 | 30.2 ± 0.2 | 30.6 ± 0.2 | 0.0 | 26.7 ± 0.5 | 9.5 | 26.7 ± 2.5 | 29.4 | 25.8 ± 0.6 | 18.8 | 33.5 ± 1.6 | 3.3 | 23.3 ± 0.4 | 77.3 |
| TNF-α | 23 ± 0.5 | 22.9 ± 0.2 | 1.2 | 18.8 ± 0.45 | 15.3 | 19.7 ± 2.5 | 25.36 | 18.5 ± 0.4 | 19.6 | 21.9 ± 0.3 | 2.2 | 17.3 ± 0.2 | 32.3 |
| CCL3 | 19 ± 0.4 | 18.9 ± 1.2 | 1.5 | 16 ± 0.4 | 6.8 | 17.4 ± 2.4 | 7.8 | 16.1 ± 0.7 | 6.9 | 18.2 ± 0.5 | 1.8 | 16.4 ± 0.5 | 3.8 |
| VCAM-1 | 29.6 ± 1.4 | 29.6 ± 1 | 1.2 | 27.3 ± 1 | 3.8 | 28 ± 2 | 6.5 | 26.9 ± 1 | 5.3 | 29 ± 1 | 1.6 | 25.4 ± 0.5 | 9.8 |
| ICAM-1 | 23.6 ± 0.7 | 23.5 ± 0.2 | 1.2 | 21.6 ± 0.5 | 3 | 23 ± 2.5 | 3.9 | 21.4 ± 0.6 | 3.9 | 23 ± 0.3 | 1.6 | 21.4 ± 0.4 | 3 |
| . | Unstimulated: Ct . | IL-36Ra . | IL-36α . | IL-36β . | IL-36γ . | IL-1β . | LPS . | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Ct . | Fold . | Ct . | Fold . | Ct . | Fold . | Ct . | Fold . | Ct . | Fold . | Ct . | Fold . | ||
| IL-6 | 29.7 ± 1.8 | 29.9 ± 1.3 | 1.0 | 21.3 ± 0.8 | 218.3 | 21.6 ± 2 | 560 | 21 ± 0.8 | 311.5 | 27.8 ± 1 | 3.3 | 18.5 ± 0.4 | 1216.2 |
| IL-12p40 | 31.1 ± 0.9 | 31.5 ± 0.8 | 0.0 | 25 ± 1.4 | 139.6 | 23.6 ± 1.5 | 426.7 | 22.8 ± 2.1 | 388 | 30 ± 1.3 | 2.5 | 20 ± 1.2 | 1638.8 |
| COX-2 | 29.5 ± 0.2 | 29.8 ± 0.3 | 1 | 22.1 ± 0.5 | 146.5 | 22.6 ± 2.7 | 322 | 21.6 ± 0.6 | 238.7 | 28 ± 0.1 | 2.9 | 20.4 ± 0.4 | 384.5 |
| CXCL1 | 28.5 ± 0.6 | 28.5 ± 0.4 | 1.1 | 20.5 ± 0.5 | 208.9 | 19.9 ± 0.3 | 320.3 | 20.4 ± 0.5 | 239.2 | 25.6 ± 0.3 | 7 | 19.6 ± 0.6 | 297 |
| CCL1 | 33.3 ± 0.9 | 33.1 ± 0.8 | 1.4 | 27.6 ± 2.4 | 68.4 | 27 ± 0.8 | 270.2 | 26 ± 2.2 | 175.4 | 31.4 ± 1.5 | 4.2 | 26.1 ± 1.3 | 107.6 |
| IL-12p35 | 34.3 ± 1.2 | 34.2 ± 0.8 | 1.5 | 27.8 ± 0.9 | 86 | 28 ± 1.7 | 216.5 | 27.4 ± 0.9 | 129.5 | 32.7 ± 1.3 | 4.5 | 27 ± 0.6 | 116.5 |
| IL-1β | 25.1 ± 0.9 | 25.2 ± 0.6 | 1.1 | 17.7 ± 0.6 | 139 | 18.7 ± 2.3 | 211.3 | 17.5 ± 0.6 | 171.6 | 23 ± 0.75 | 5 | 16.9 ± 0.5 | 185 |
| IL-23p19 | 30 ± 1.1 | 30.3 ± 0.9 | 1 | 24 ± 1.2 | 53 | 22.2 ± 0.9 | 207.4 | 23.2 ± 1 | 97.2 | 28.5 ± 0.7 | 2.9 | 22.4 ± 1 | 134.8 |
| GM-CSF | 33 ± 0.7 | 32.6 ± 0.9 | 2 | 27.6 ± 0.8 | 40.6 | 27.8 ± 2 | 111.2 | 26.5 ± 0.7 | 88.2 | 29.5 ± 0.6 | 13.4 | 29 ± 0.5 | 11.8 |
| IL-10 | 34.5 ± 1 | 40 ± 0.0 | 0 | 31.2 ± 0.5 | 16 | 30.4 ± 2.8 | 91 | 29.7 ± 0.7 | 49 | 34.9 ± 0.3 | 2 | 29.5 ± 0.1 | 42 |
| CXCL10 | 28 ± 1.7 | 28 ± 1.5 | 1.5 | 25.8 ± 1.4 | 3.9 | 25.5 ± 4.3 | 47 | 25.5 ± 1.4 | 5 | 27.1 ± 1 | 2 | 18.25 ± 0.7 | 644.3 |
| NOS2 | 30.2 ± 0.2 | 30.6 ± 0.2 | 0.0 | 26.7 ± 0.5 | 9.5 | 26.7 ± 2.5 | 29.4 | 25.8 ± 0.6 | 18.8 | 33.5 ± 1.6 | 3.3 | 23.3 ± 0.4 | 77.3 |
| TNF-α | 23 ± 0.5 | 22.9 ± 0.2 | 1.2 | 18.8 ± 0.45 | 15.3 | 19.7 ± 2.5 | 25.36 | 18.5 ± 0.4 | 19.6 | 21.9 ± 0.3 | 2.2 | 17.3 ± 0.2 | 32.3 |
| CCL3 | 19 ± 0.4 | 18.9 ± 1.2 | 1.5 | 16 ± 0.4 | 6.8 | 17.4 ± 2.4 | 7.8 | 16.1 ± 0.7 | 6.9 | 18.2 ± 0.5 | 1.8 | 16.4 ± 0.5 | 3.8 |
| VCAM-1 | 29.6 ± 1.4 | 29.6 ± 1 | 1.2 | 27.3 ± 1 | 3.8 | 28 ± 2 | 6.5 | 26.9 ± 1 | 5.3 | 29 ± 1 | 1.6 | 25.4 ± 0.5 | 9.8 |
| ICAM-1 | 23.6 ± 0.7 | 23.5 ± 0.2 | 1.2 | 21.6 ± 0.5 | 3 | 23 ± 2.5 | 3.9 | 21.4 ± 0.6 | 3.9 | 23 ± 0.3 | 1.6 | 21.4 ± 0.4 | 3 |
BMDCs from WT mice were stimulated or not with IL-36Ra, IL-36α, IL-36β, IL-36γ, IL-1β, or LPS at a concentration of 100 ng/mL. Two hours after stimulation, mRNA was extracted and analyzed for mRNA expression using TaqMan low density array. The threshold cycle values (Ct) of each transcript represent the mean ± SD of triplicates in the same experiment. Hypoxanthine guanine phosphoribosyl transferase (HPRT) gene expression was assessed as an endogenous reference and used for fold increase normalization. The stimulatory activities of IL-36R ligands, IL-1β, or LPS were measured in fold increase compared with the nonstimulated cells. Data show only cytokines, chemokines, or growth factors significantly induced (P < .05) by the different stimuli compared with unstimulated cells. Results are representative of 3 independent experiments with similar results.