Fig 1: Elevated IKKα levels in inflammatory mice ears, evoked by IL-17 and IL-22 injection. Skin inflammation, edema, and acanthosis, which were involved in the infiltration of neutrophils and dendritic cells into the ear skin of C57BL/6 WT mice, but less profound inflammatory changes, were noted in the sham controls. Each ear of C57BL/6 WT mice were injected intradermally with 3µg of IL-17 and IL-22 for two consecutive days in a total volume of 30µl. The control group of mice was injected with the same volumes of phosphate-buffered saline (PBS) via an identical route.
Fig 2: Both IL-17 and IL-22 induce an increase in IKKα expression in Neoderm. Neoderm tissues were cultured under 100 ng/ml of IL-17 or IL-22 for 48 hours and fixed for H&E and immunohistochemical staining assays. The results shown are representative of the three independent batches of Neoderm experiments.
Fig 3: Depletion of Tregs Induces a Mild Systemic Inflammatory Response in the Absence of Intra-Amniotic Inflammation(A–H) Foxp3DTR dams underwent partial or total Treg depletion. Controls were injected with sterile 13 PBS. Mice were euthanized approximately 4 h after the second DT or PBS injection, and maternal plasma samples and amniotic fluid were collected. Concentrations of (A) IL-6, (B) CCL2, (C) IL-1ß, (D) TNF-a, (E) IFN?, (F) CCL7, (G) IL-22, and (H) IL-10 in the maternal plasma (n = 7 per group). Data are shown as medians with interquartile ranges and minimum/maximum ranges. C57BL/6 dams were intravenously injected with recombinant mouse IFN? (1.4 pg/100 µL), CCL7 (218 pg/100 µL), IL-22 (48 pg/100 µL), or a combination of all three. Controls were injected with 100 µL of sterile 13 PBS alone.(I) Gestational length of cytokine-injected dams. Data are shown as means with standard deviations (n = 3–6 per group).(J) Rate of early-term or full-term delivery of cytokine-injected dams (n = 3–6 per group).(K) Percentage of survival from birth until 3 weeks postpartum for neonates born to cytokine-injected dams (n = 3–6 litters per group).(L–S) Concentrations of (L) IL-6, (M) CXCL10, (N) CCL2, (O) IL-1ß, (P) TNF-a, (Q) IFN?, (R) IL-10, and (S) IL-4 in the amniotic fluid (n = 5–7 per group). Data are shown as medians with interquartile ranges and minimum/maximum ranges.Statistical analysis was performed using the Mantel-Cox test for survival curves, and Kruskal-Wallis or ANOVA tests with correction for multiple comparisons. See also Figure S5.
Fig 4: The effects of IL-17 and IL-22 treatment on the expression of IKKα and phosphorylated IκB in HaCaT cells. Cells were treated with IL-17 and IL-22 up to a concentration of 100 ng/ml in a calcium concentration of 2.8 mM and cultured for 24 hours. (A) The immunoblot shown is representative of three independent experiments. (B) The pixel densities are divided by β-actin densities to correct the values, and the data are expressed as the means±standard error of the mean at three independent experiments. *p<0.05.
Fig 5: IL-11 induces signals to colonic epithelial cells and fibroblasts.a Colonic epithelial organoids and fibroblasts were established from three independent wild-type mice as described in the methods. Expressions of the indicated genes were determined using qPCR. Results are mean ± SEM (n = 3 mice). Results are representative of two independent experiments. b Colonic epithelial organoids and fibroblasts were unstimulated or stimulated for 30 min with IL-11 (10 or 100 ng/mL) or IL-22 (10 or 100 ng/mL). Total STAT3 and phosphorylated STAT3 (pSTAT3) were analyzed by Western blotting. STAT3 and pSTAT3 signaling intensities were calculated by Fiji, and the relative ratio of pSTAT3/STAT3 is shown. Results are representative of two independent experiments (a, b). c, d Relative expression of Il11ra1 and Il6st in colon organoids. Colon organoids were established from WT and Apcdelta716 mice carrying a truncated Apc gene. Expression of the indicated genes in WT and Apcdelta716 organoids was determined using qPCR (c). Results are mean ± SEM (n = 3 independently established organoids from different mice). Colon organoids were not stimulated or stimulated with IL-11 (100 ng/mL) for 30 min (d). Total STAT3, phosphorylated STAT3 (pSTAT3), total ERK, and phosphorylated ERK (pERK) were analyzed and described as in (b). Results are representative of two independent experiments. e, f Administration of IL-11R agonist induces expression of the genes expressed in IL-11+ cells. We injected 8-week-old wild-type mice with 10 µg IL-11R agonist. At 3 h after injection, mRNA was isolated from the whole colon, and gene expression was analyzed using microarray analysis as described in “Methods” (n = 2 for untreated samples; n = 3 for injected samples). Volcano plot of whole genes (e). The horizontal line indicates differentially regulated genes in the whole colon after IL-11R agonist injection compared with the whole colon before injection, shown in log2. The vertical line indicates P values, shown in -log10. Red dots indicate significantly upregulated genes. Venn diagram of genes enriched in FACS-sorted IL-11+ fibroblasts and genes with elevated expression in the whole colon after IL-11R agonist treatment compared with an untreated colon (f). Nineteen overlapping genes are shown. g Venn diagram of genes enriched in FACS-sorted IL-11+ fibroblasts and genes induced by IL-11 stimulation in ColVI+ fibroblasts. Five overlapping genes are shown. Statistical significance was determined using the two-tailed unpaired Student’s t-test (a, c). p-values were calculated by two-tailed local-pooled-error test (e). Source data are provided as a Source Data file.
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