Journal: Journal of Translational Medicine

Article Title: IL20RB signaling enhances stemness and chemotherapy resistance in pancreatic cancer

doi: 10.1186/s12967-023-04800-5

Figure Lengend Snippet: IL20RB is highly expressed in tumor tissues of pancreatic cancer patients and is associated with poor prognosis. A Representative immunofluorescence images of IL20RB and pan-CK in human pancreatic cancer sections and surrounding tissues, scale bar = 40 µm. B Statistical analysis of IL20RB staining intensity in pancreatic cancer tissue sections from patients at different stages. C Correlation analysis between IL20RB staining intensity and overall survival of patients with pancreatic cancer, and the cut-off value was the median of IL20RB staining intensity. D IL20RB mRNA expression in pan-cancer and normal tissues, and the red arrow indicates pancreatic cancer, data from TCGA. E IL20RB mRNA expression in normal pancreatic tissues and pancreatic cancer tissues, data from TCGA. F mRNA expression of IL20RB in pancreatic cancer and paired adjacent tissues, data from Gene Expression Omnibus (GSE15471). G Correlation analysis of IL20RB mRNA expression level and overall survival of patients with pancreatic cancer, data from TIMER2.0 database. ( H–K ) Correlation analysis of IL20RB mRNA expression at T stage, N stage and AJCC stage and survival status of patients with pancreatic cancer, data from TCGA

Article Snippet: Antibodies: rabbit anti-IL20RB antibody (dil. 1:50, Proteintech, 20,521-1-AP); rabbit anti-NANOG antibody (dil. 1:500, Proteintech, 14,295-1-AP); rabbit anti-SOX2 antibody (dil. 1:150, Proteintech, 11,064-1-AP); rabbit phospho-Stat3 (Tyr705) antibody (dil. 1:300, Cell Signaling Technology, 9145); mouse anti-pan-CK antibody (dil. 1:500, Abcam, ab215838); rabbit anti-IL-19 antibody (dil. 1:500, Bioss, bs-10087R) (dil., dilution).

Techniques: Immunofluorescence, Staining, Expressing, Gene Expression

Journal: Journal of Translational Medicine

Article Title: IL20RB signaling enhances stemness and chemotherapy resistance in pancreatic cancer

doi: 10.1186/s12967-023-04800-5

Figure Lengend Snippet: IL20RB promotes stemness and chemoresistance in pancreatic cancer in vitro. A Real-time quantitative PCR analysis of IL20RB mRNA expression in pancreatic cancer cell lines PANC-1 and MIA PaCa-2 (n = 3 in each group). B Western blot analysis of IL20RB protein expression in PANC-1 and MIA PaCa-2 (n = 3 in each group). C – D Sphere-forming assay and statistical analysis of PANC-1 and MIA PaCa-2 (n = 3 per group). E – F Flow cytometry and statistical analyses of side population cells in PANC-1 and MIA PaCa-2 (n = 3 in each group). G Real-time quantitative PCR analysis of IL20RB, NANOG, SOX2 and POU5F1 mRNA expression in PANC-1 and MIA PaCa-2 (n = 3 in each group). H Western blot analysis of IL20RB, NANOG and SOX2 protein expression in PANC-1 and MIA PaCa-2 (n = 3 in each group). I – J Results of colony formation assay and statistical analysis of pancreatic cancer cell lines treated with 10 nM gemcitabine for 24 h. K – L Representative immunofluorescence images and statistical analysis of IL20RB, NANOG and SOX2 in human pancreatic cancer tissue sections, scale bar = 40 µm

Article Snippet: Antibodies: rabbit anti-IL20RB antibody (dil. 1:50, Proteintech, 20,521-1-AP); rabbit anti-NANOG antibody (dil. 1:500, Proteintech, 14,295-1-AP); rabbit anti-SOX2 antibody (dil. 1:150, Proteintech, 11,064-1-AP); rabbit phospho-Stat3 (Tyr705) antibody (dil. 1:300, Cell Signaling Technology, 9145); mouse anti-pan-CK antibody (dil. 1:500, Abcam, ab215838); rabbit anti-IL-19 antibody (dil. 1:500, Bioss, bs-10087R) (dil., dilution).

Techniques: In Vitro, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Flow Cytometry, Colony Assay, Immunofluorescence

Journal: Journal of Translational Medicine

Article Title: IL20RB signaling enhances stemness and chemotherapy resistance in pancreatic cancer

doi: 10.1186/s12967-023-04800-5

Figure Lengend Snippet: IL20RB promotes stemness and chemoresistance in pancreatic cancer in vivo. A – B Tumor formation results after subcutaneous injection of 10 5 , 10 6 and 10 7 PANC-1, respectively, into nude mice. C Tumor growth curve (n = 5 per group). D Statistical analysis of tumor weight at the end of the trial (n = 5 per group). E – F Representative immunohistochemical (IHC) images of NANOG and SOX2 in tumor tissue sections and statistical analysis of IHC scores, scale bar = 40 µm. G Western blot analysis of NANOG, SOX2 and IL20RB in tumor tissues. ( H ) 10^7 PANC-1 was injected subcutaneously into nude mice, and then gemcitabine 100 mg/kg was intraperitoneally injected twice a week for 4 weeks. I Statistical analysis of tumor weight at the endpoint. J Tumor growth curve. K – L Representative IHC images of pan-CK in tumor tissue sections and statistical analysis of IHC scores, scale bar = 40 µm

Article Snippet: Antibodies: rabbit anti-IL20RB antibody (dil. 1:50, Proteintech, 20,521-1-AP); rabbit anti-NANOG antibody (dil. 1:500, Proteintech, 14,295-1-AP); rabbit anti-SOX2 antibody (dil. 1:150, Proteintech, 11,064-1-AP); rabbit phospho-Stat3 (Tyr705) antibody (dil. 1:300, Cell Signaling Technology, 9145); mouse anti-pan-CK antibody (dil. 1:500, Abcam, ab215838); rabbit anti-IL-19 antibody (dil. 1:500, Bioss, bs-10087R) (dil., dilution).

Techniques: In Vivo, Injection, Immunohistochemical staining, Western Blot

Journal: Journal of Translational Medicine

Article Title: IL20RB signaling enhances stemness and chemotherapy resistance in pancreatic cancer

doi: 10.1186/s12967-023-04800-5

Figure Lengend Snippet: IL20RB activates STAT3 phosphorylation to promote pancreatic cancer stemness and chemotherapy resistance. A Western blot analysis of IL20RB, STAT3 and p-STAT3 (Tyr705) proteins in pancreatic cancer cell line MIA PaCa-2. B Western blot analysis of IL20RB, STAT3 and p-STAT3 (Tyr705) in pancreatic cancer cell line PANC-1. C Real-time quantitative PCR analysis of the mRNA expression of IL20RB, NANOG, SOX2 and POU5F1 in MIA PaCa-2 treated with the vehicle or STAT3 inhibitor (Stattic), respectively (n = 3 in each group). ( D ) Western blot analysis of IL20RB, STAT3, p-STAT3 (Tyr705), NANOG and SOX2 proteins in MIA PaCa-2 after the vehicle or Stattic treatment. E Sphere-forming assay and statistical analysis of MIA PaCa-2 after treatment with the vehicle or Stattic (n = 3 per group). F Colony formation assay and statistical analysis of MIA PaCa-2 treated with 10 nM gemcitabine for 24 h with or without Stattic (n = 3 per group). G Representative immunofluorescence images of IL20RB, pan-CK and p-STAT3 in human pancreatic cancer tissue sections, scale bar = 40 µm

Article Snippet: Antibodies: rabbit anti-IL20RB antibody (dil. 1:50, Proteintech, 20,521-1-AP); rabbit anti-NANOG antibody (dil. 1:500, Proteintech, 14,295-1-AP); rabbit anti-SOX2 antibody (dil. 1:150, Proteintech, 11,064-1-AP); rabbit phospho-Stat3 (Tyr705) antibody (dil. 1:300, Cell Signaling Technology, 9145); mouse anti-pan-CK antibody (dil. 1:500, Abcam, ab215838); rabbit anti-IL-19 antibody (dil. 1:500, Bioss, bs-10087R) (dil., dilution).

Techniques: Phospho-proteomics, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Colony Assay, Immunofluorescence

Journal: Journal of Translational Medicine

Article Title: IL20RB signaling enhances stemness and chemotherapy resistance in pancreatic cancer

doi: 10.1186/s12967-023-04800-5

Figure Lengend Snippet: IL-19 derived from the microenvironment activates the IL20RB-STAT3 pathway to promote stemness and chemoresistance in pancreatic cancer. A Flow cytometry analysis of side population cells in the pancreatic cancer cell line MIA PaCa-2 after treatment with the ligands of IL20RB (recombinant IL-19, IL-20 and IL-24 proteins) for 24 h and statistical analysis (n = 3 per group). B Real-time quantitative PCR analysis of the NANOG, SOX2 and POU5F1 mRNA expression in pancreatic cancer cell line PANC-1 after treatment with recombinant IL-19, IL-20 and IL-24 proteins for 24 h (n = 3 in each group). C Western blot analysis of NANOG, SOX2, STAT3, and p-STAT3 (Tyr705) proteins in PANC-1 after treatment with recombinant IL-19, IL-20 and IL-24 proteins for 24 h, respectively. ( D ) Real-time quantitative PCR analysis of the NANOG, SOX2 and POU5F1 mRNA expression in PANC-1 after treatment with 0, 20, 40 and 60 ng/mL recombinant IL-19 protein for 24 h (n = 3 per group). E – F Results of Western blot analysis of NANOG, SOX2, STAT3, p-STAT3 (Tyr705) and IL20RB proteins in PANC-1 and MIA PaCa-2 after treatment with the vehicle or recombinant IL-19 protein for 24 h. G Sphere-forming assay and statistical analysis of PANC-1 treated with the vehicle or recombinant IL-19 protein for 24 h (n = 3 per group). H Western blot analysis of NANOG, SOX2, STAT3 and p-STAT3 (Tyr705) proteins in PANC-1 after treatment with the vehicle, recombinant IL-19 protein, or Stattic for 24 h. I Results of colony formation assay and statistical analysis of the MIA PaCa-2 after treatment with 10 nM gemcitabine for 24 h, followed by treatment with the vehicle, recombinant IL-19 protein alone or in combination with Stattic (n = 3 per group). J Representative immunofluorescence images of IL-19 and pan-CK in human pancreatic cancer tissue sections, scale bar = 40 µm

Article Snippet: Antibodies: rabbit anti-IL20RB antibody (dil. 1:50, Proteintech, 20,521-1-AP); rabbit anti-NANOG antibody (dil. 1:500, Proteintech, 14,295-1-AP); rabbit anti-SOX2 antibody (dil. 1:150, Proteintech, 11,064-1-AP); rabbit phospho-Stat3 (Tyr705) antibody (dil. 1:300, Cell Signaling Technology, 9145); mouse anti-pan-CK antibody (dil. 1:500, Abcam, ab215838); rabbit anti-IL-19 antibody (dil. 1:500, Bioss, bs-10087R) (dil., dilution).

Techniques: Derivative Assay, Flow Cytometry, Recombinant, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Colony Assay, Immunofluorescence