Journal: International Journal of Molecular Sciences

Article Title: The Microarchitecture of Pancreatic Cancer as Measured by Diffusion-Weighted Magnetic Resonance Imaging Is Altered by T Cells with a Tumor Promoting Th17 Phenotype

doi: 10.3390/ijms21010346

Figure Lengend Snippet: Receptors for IL21 and IL26 in PDAC. ( A ) Immunohistochemistry (IHC) of PDAC samples with no expression and high expression of IL10RB, IL20RA, or IL21R, in comparison to non-neoplastic pancreatic parenchyma. Arrow head: tumor cells ( B ) Double staining for IL10RB (red) and IL20RA (brown) shows co-localization (arrow, red-brown cells). ( C ) Bubble charts show the dependencies of the Allred score for IL20RA from Allred IL10RB (left), Allred IL21R from Allred IL10RB (middle), and Allred IL21R from Allred IL20RA (right). The diameter of the bubbles indicates the frequency of occurrence.

Article Snippet: After 24 h, cells were fixed with 2 % PFA and incubated with a rabbit anti-human IL10RB antibody (dilution: 1:50; NBP2–33492, NovusBiologicals) and a mouse anti-human IL20RA antibody (dilution: 1:50; NBP2–11695, Novus Biologicals), at 4 °C overnight.

Techniques: Immunohistochemistry, Expressing, Comparison, Double Staining

Journal: International Journal of Molecular Sciences

Article Title: The Microarchitecture of Pancreatic Cancer as Measured by Diffusion-Weighted Magnetic Resonance Imaging Is Altered by T Cells with a Tumor Promoting Th17 Phenotype

doi: 10.3390/ijms21010346

Figure Lengend Snippet: PDAC cell lines. ( A ) Laser scan microscopy and Western blotting of the PDAC cell lines AsPC-1, BxPC-3, and Panc-1 incubated with anti-IL10RB and anti-IL20RA antibodies shows expression and colocalization of both receptors in all three cell lines. ( B ) Western blotting of PDAC cells shows IL26 induced phosphorylation of STAT3 (left panel) and ERK1/2 (right panel) in a time-dependent manner.

Article Snippet: After 24 h, cells were fixed with 2 % PFA and incubated with a rabbit anti-human IL10RB antibody (dilution: 1:50; NBP2–33492, NovusBiologicals) and a mouse anti-human IL20RA antibody (dilution: 1:50; NBP2–11695, Novus Biologicals), at 4 °C overnight.

Techniques: Microscopy, Western Blot, Incubation, Expressing, Phospho-proteomics

Journal: Frontiers in Oncology

Article Title: Super-Enhancer Induced IL-20RA Promotes Proliferation/Metastasis and Immune Evasion in Colorectal Cancer

doi: 10.3389/fonc.2021.724655

Figure Lengend Snippet: IL-20RA was activated by super-enhancers in colorectal cancer. (A) Genome-browser screenshot showing the binding of H3K27ac to the regulatory region of IL-20RA in primary CRC tumor cells, cell lines and normal colon tissues. (B) Super-enhancer inhibition decreased the cell growth of colorectal cancer cell lines. The viabilities of RKO, HT-29, and LoVo cells were detected after 0, 24, 48, and 72 hours of JQ-1 or i-BET treatment. (C) Super-enhancer inhibition decreased the expression of IL-20RA in CRC cells. The expression of IL-20RA was examined in RKO, HT-29, and LoVo cells after 48 hours of JQ-1 or i-BET treatment. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Primary antibodies included IL-20RA (mouse mAb, #sc-80065, 1:500; Santa Cruz, CA, USA), GAPDH (rabbit mAb, #5174, 1:5000; CST, Danvers, MA, USA), Snail (rabbit mAb, #3879, 1:1000; CST), and Slug (rabbit mAb, #9585, 1:1000; CST).

Techniques: Binding Assay, Inhibition, Expressing

Journal: Frontiers in Oncology

Article Title: Super-Enhancer Induced IL-20RA Promotes Proliferation/Metastasis and Immune Evasion in Colorectal Cancer

doi: 10.3389/fonc.2021.724655

Figure Lengend Snippet: IL-20RA was highly expressed in colorectal cancer. (A) IHC staining images of IL-20RA between colorectal cancer tissues and paired normal tissues in the microarray. (B) Statistical analysis of the IHC scores of IL-20RA in colorectal cancer and paired normal tissues of 118 human colorectal cancer patients in different UICC stages. (25 for stage 1; 42 for stage 2; 40 for stage 3; 11 for stage 4). (C) IL-20RA was highly expressed in CRC compared to normal colon tissues. The expression of IL-20RA in CRC and normal colon tissues was retrieved from the RNA-seq data of the TCGA-COAD cohort. (D) IL-20RA levels in plasma between 52 colorectal cases and 17 normal persons (10 for stage 1; 16 for stage 2; 18 for stage 3; 8 for stage 4). *P < 0.05, **P < 0.01, ****P < 0.0001.

Article Snippet: Primary antibodies included IL-20RA (mouse mAb, #sc-80065, 1:500; Santa Cruz, CA, USA), GAPDH (rabbit mAb, #5174, 1:5000; CST, Danvers, MA, USA), Snail (rabbit mAb, #3879, 1:1000; CST), and Slug (rabbit mAb, #9585, 1:1000; CST).

Techniques: Immunohistochemistry, Microarray, Expressing, RNA Sequencing, Clinical Proteomics

Journal: Frontiers in Oncology

Article Title: Super-Enhancer Induced IL-20RA Promotes Proliferation/Metastasis and Immune Evasion in Colorectal Cancer

doi: 10.3389/fonc.2021.724655

Figure Lengend Snippet: Correlation between IL-20RA expression and clinical characteristics in 118 colorectal cancer tissues.

Article Snippet: Primary antibodies included IL-20RA (mouse mAb, #sc-80065, 1:500; Santa Cruz, CA, USA), GAPDH (rabbit mAb, #5174, 1:5000; CST, Danvers, MA, USA), Snail (rabbit mAb, #3879, 1:1000; CST), and Slug (rabbit mAb, #9585, 1:1000; CST).

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Super-Enhancer Induced IL-20RA Promotes Proliferation/Metastasis and Immune Evasion in Colorectal Cancer

doi: 10.3389/fonc.2021.724655

Figure Lengend Snippet: IL-20RA was associated with oncogenic and immune pathways. (A, B) Differently expressed genes showed by volcano plot using TCGA-COAD datasets and RNA-seq. (C) IL-20RA-related pathways were analyzed with Gene set variation analysis. (D) Heatmap showed genes differentially expressed in LoVo cells with or without IL-20RA knockdown. (E, F) Gene ontology analysis of genes down- (E) and up-regulated (F) post-IL-20RA knockdown. (G) Expression of IL-20RA, Snail and Slug at the protein level in shIL-20RA LoVo cells detected by immunoblotting. The data were obtained from 3 independent experiments and the presented as the mean ± SD (GAPDH was used as a loading control).

Article Snippet: Primary antibodies included IL-20RA (mouse mAb, #sc-80065, 1:500; Santa Cruz, CA, USA), GAPDH (rabbit mAb, #5174, 1:5000; CST, Danvers, MA, USA), Snail (rabbit mAb, #3879, 1:1000; CST), and Slug (rabbit mAb, #9585, 1:1000; CST).

Techniques: RNA Sequencing, Knockdown, Expressing, Western Blot, Control

Journal: Frontiers in Oncology

Article Title: Super-Enhancer Induced IL-20RA Promotes Proliferation/Metastasis and Immune Evasion in Colorectal Cancer

doi: 10.3389/fonc.2021.724655

Figure Lengend Snippet: Knockdown of IL-20RA inhibited the growth and metastasis of colorectal cancer cells. (A) Cell proliferation of shIL-20RA was decreased in LoVo cells. Cell viability was detected at 0, 24, 48, and 72 hours by CCK-8 assay. (B, C) Wound healing assay of shIL-20RA in LoVo cells. Healing ability was decreased with shIL-20RA. The images were photographed at 0, 24, and 48 hours. (D, E) The migration ability of LoVo cells treated with shIL-20RA was decreased as shown by Transwell analyses. The number of migrated cells was counted under the average of 5 random fields. (F, G) The invasion ability of LoVo cells treated with shIL-20RA was decreased as shown by Transwell analyses. The number of invasive cells was counted under the average of 5 random fields. ****P < 0.0001.

Article Snippet: Primary antibodies included IL-20RA (mouse mAb, #sc-80065, 1:500; Santa Cruz, CA, USA), GAPDH (rabbit mAb, #5174, 1:5000; CST, Danvers, MA, USA), Snail (rabbit mAb, #3879, 1:1000; CST), and Slug (rabbit mAb, #9585, 1:1000; CST).

Techniques: Knockdown, CCK-8 Assay, Wound Healing Assay, Migration

Journal: Frontiers in Oncology

Article Title: Super-Enhancer Induced IL-20RA Promotes Proliferation/Metastasis and Immune Evasion in Colorectal Cancer

doi: 10.3389/fonc.2021.724655

Figure Lengend Snippet: IL-20RA was related to infiltration level of immune cells. (A, B) Heatmap and boxplot derived from gene set variation analysis showing varied levels of infiltrating immune cells between high- and low-IL-20RA groups. (C-E) Gene set enrichment analysis of genes related to IFN-γ response (C) , IL-6-Jak-STAT3 (D) and adaptive immune response (E) in genes positively or negatively correlated with IL-20RA expression. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, NS, no significance.

Article Snippet: Primary antibodies included IL-20RA (mouse mAb, #sc-80065, 1:500; Santa Cruz, CA, USA), GAPDH (rabbit mAb, #5174, 1:5000; CST, Danvers, MA, USA), Snail (rabbit mAb, #3879, 1:1000; CST), and Slug (rabbit mAb, #9585, 1:1000; CST).

Techniques: Derivative Assay, Expressing

Journal: International Journal of Molecular Sciences

Article Title: The Microarchitecture of Pancreatic Cancer as Measured by Diffusion-Weighted Magnetic Resonance Imaging Is Altered by T Cells with a Tumor Promoting Th17 Phenotype

doi: 10.3390/ijms21010346

Figure Lengend Snippet: Receptors for IL21 and IL26 in PDAC. ( A ) Immunohistochemistry (IHC) of PDAC samples with no expression and high expression of IL10RB, IL20RA, or IL21R, in comparison to non-neoplastic pancreatic parenchyma. Arrow head: tumor cells ( B ) Double staining for IL10RB (red) and IL20RA (brown) shows co-localization (arrow, red-brown cells). ( C ) Bubble charts show the dependencies of the Allred score for IL20RA from Allred IL10RB (left), Allred IL21R from Allred IL10RB (middle), and Allred IL21R from Allred IL20RA (right). The diameter of the bubbles indicates the frequency of occurrence.

Article Snippet: The following primary anti-human antibodies were used: rabbit anti-human IL10RB (dilution: 1:400; NovusBiologicals), mouse anti-human IL20RA (dilution: 1:400; Novus Biologicals), rabbit anti-human ERK (dilution: 1:1000; Cell Signaling Technology), rabbit anti-human pERK (dilution: 1:1000; Cell Signaling Technology), rabbit anti-human STAT3 (dilution: 1:1000; Cell Signaling Technology), rabbit anti-human pSTAT3 (dilution: 1:1000; Cell Signaling Technology), mouse anti-human β -actin (dilution: 1:10000; MP Biomedicals, Eschwege, Germany) and mouse anti-human β -tubulin (dilution: 1:300; Santa Cruz Biotechnologies).

Techniques: Immunohistochemistry, Expressing, Comparison, Double Staining

Journal: International Journal of Molecular Sciences

Article Title: The Microarchitecture of Pancreatic Cancer as Measured by Diffusion-Weighted Magnetic Resonance Imaging Is Altered by T Cells with a Tumor Promoting Th17 Phenotype

doi: 10.3390/ijms21010346

Figure Lengend Snippet: PDAC cell lines. ( A ) Laser scan microscopy and Western blotting of the PDAC cell lines AsPC-1, BxPC-3, and Panc-1 incubated with anti-IL10RB and anti-IL20RA antibodies shows expression and colocalization of both receptors in all three cell lines. ( B ) Western blotting of PDAC cells shows IL26 induced phosphorylation of STAT3 (left panel) and ERK1/2 (right panel) in a time-dependent manner.

Article Snippet: The following primary anti-human antibodies were used: rabbit anti-human IL10RB (dilution: 1:400; NovusBiologicals), mouse anti-human IL20RA (dilution: 1:400; Novus Biologicals), rabbit anti-human ERK (dilution: 1:1000; Cell Signaling Technology), rabbit anti-human pERK (dilution: 1:1000; Cell Signaling Technology), rabbit anti-human STAT3 (dilution: 1:1000; Cell Signaling Technology), rabbit anti-human pSTAT3 (dilution: 1:1000; Cell Signaling Technology), mouse anti-human β -actin (dilution: 1:10000; MP Biomedicals, Eschwege, Germany) and mouse anti-human β -tubulin (dilution: 1:300; Santa Cruz Biotechnologies).

Techniques: Microscopy, Western Blot, Incubation, Expressing, Phospho-proteomics

Journal: Experimental eye research

Article Title: IL-20 Promotes Epithelial Healing of the Injured Mouse Cornea

doi: 10.1016/j.exer.2016.11.006

Figure Lengend Snippet: IL-20 and IL-20RA are detected in the mouse cornea. (A) Binding of TRITC-anti-IL-20RA or TRITC-IgG to normal mouse corneal epithelium. (B) Binding of FITC-anti-IL-20 or FITC-IgG to normal mouse corneal epithelium, and (C) to stromal keratocytes. (D) ELISA determinations of IL-20 from extracted corneas without wounding and at various times after central epithelial abrasion (n=10 corneas pooled at each timepoint; ** p<0.01). (E) Human epithelial cell line (HTCEpi cells) stained with FITC-anti-IL-20 or FITC-IgG and (F) with TRITC-anti-IL-20RA or TRITC-IgG.

Article Snippet: To evaluate IL-20 and IL20RA expression in human corneal epithelial cells, we cultured human telomerase human corneal epithelial cells (HTCEpi) for 4 hours in KBM-2 medium (Fisher Scientific, Pittsburgh, PA) containing 10 units/ml IL-1β (R&D Systems, Minneapolis, MN) after which the cells were labeled with anti-IL-20 (Abcam, Cambridge, MA) or anti-IL-20RA (Millipore, Billerica, MA) antibodies.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Staining