Journal: Journal of Personalized Medicine

Article Title: IL4Rα and IL13Rα1 Are Involved in the Development of Human Gallbladder Cancer

doi: 10.3390/jpm12020249

Figure Lengend Snippet: Immunohistochemical expression of IL4Rα and IL13Rα1 in human gallbladder carcinoma tissue. ( A ) The nuclear and cytoplasmic expression of IL4Rα and IL13Rα1 are seen in positive examples of adenocarcinoma and squamous cell carcinoma components of gallbladder carcinomas. Original magnification: x400. ( B ) Statistical analysis to determine immunohistochemical positivity of the expression of IL4Rα and IL13Rα1. The cut-off points are determined by using receiver operating characteristic curve analysis at the points with the highest area under the curve (AUC) to predict cancer related death of patients. The cut-off points for the expression of nuclear IL4Rα (Nu-IL4Rα, black arrow), cytoplasmic IL4Rα (Cy-IL4Rα, arrowhead), nuclear IL13Rα1 (Nu-IL13Rα1, empty arrow), and cytoplasmic IL13Rα1 (Cy-IL13Rα1, empty arrowhead) are indicated in the receiver operating characteristic curve.

Article Snippet: 5 uL of control, IL4Rα, or IL13Rα1 siRNA (Santa Cruz, the concentration of siRNA stock solution is 10 uM) and 15 µL of lipofectamine 2000 (Waltham, MA, USA) were added to media respectively and incubated for 15 min. After incubation, the media containing siRNA and the media containing lipofectamine 2000 were mixed and incubated for 15 min. After 15 min of incubation, cells were washed twice with PBS and the siRNA and lipofectamine 2000 mixtures were added to cells.

Techniques: Immunohistochemical staining, Expressing

Journal: Journal of Personalized Medicine

Article Title: IL4Rα and IL13Rα1 Are Involved in the Development of Human Gallbladder Cancer

doi: 10.3390/jpm12020249

Figure Lengend Snippet: Clinical variables and the expression of IL4Rα and IL13Rα1 in gallbladder carcinomas.

Article Snippet: 5 uL of control, IL4Rα, or IL13Rα1 siRNA (Santa Cruz, the concentration of siRNA stock solution is 10 uM) and 15 µL of lipofectamine 2000 (Waltham, MA, USA) were added to media respectively and incubated for 15 min. After incubation, the media containing siRNA and the media containing lipofectamine 2000 were mixed and incubated for 15 min. After 15 min of incubation, cells were washed twice with PBS and the siRNA and lipofectamine 2000 mixtures were added to cells.

Techniques: Expressing

Journal: Journal of Personalized Medicine

Article Title: IL4Rα and IL13Rα1 Are Involved in the Development of Human Gallbladder Cancer

doi: 10.3390/jpm12020249

Figure Lengend Snippet: Univariate analysis of overall survival and relapse-free survival in gallbladder carcinoma patients.

Article Snippet: 5 uL of control, IL4Rα, or IL13Rα1 siRNA (Santa Cruz, the concentration of siRNA stock solution is 10 uM) and 15 µL of lipofectamine 2000 (Waltham, MA, USA) were added to media respectively and incubated for 15 min. After incubation, the media containing siRNA and the media containing lipofectamine 2000 were mixed and incubated for 15 min. After 15 min of incubation, cells were washed twice with PBS and the siRNA and lipofectamine 2000 mixtures were added to cells.

Techniques:

Journal: Journal of Personalized Medicine

Article Title: IL4Rα and IL13Rα1 Are Involved in the Development of Human Gallbladder Cancer

doi: 10.3390/jpm12020249

Figure Lengend Snippet: Survival analysis according to the expression of IL4Rα and IL13Rα1 in gallbladder carcinoma patients. Kaplan–Meier survival curves of overall survival and relapse-free survival according to the expression of nuclear IL4Rα (Nu-IL4Rα) ( A ), cytoplasmic IL4Rα (Cy-cIL4Rα) ( B ), nuclear IL13Rα1 (Nu-IL13Rα1) ( C ), and cytoplasmic IL13Rα1 (Cy-IL13Rα1) ( D ).

Article Snippet: 5 uL of control, IL4Rα, or IL13Rα1 siRNA (Santa Cruz, the concentration of siRNA stock solution is 10 uM) and 15 µL of lipofectamine 2000 (Waltham, MA, USA) were added to media respectively and incubated for 15 min. After incubation, the media containing siRNA and the media containing lipofectamine 2000 were mixed and incubated for 15 min. After 15 min of incubation, cells were washed twice with PBS and the siRNA and lipofectamine 2000 mixtures were added to cells.

Techniques: Expressing

Journal: Journal of Personalized Medicine

Article Title: IL4Rα and IL13Rα1 Are Involved in the Development of Human Gallbladder Cancer

doi: 10.3390/jpm12020249

Figure Lengend Snippet: Anti-proliferative effect by knockdown of IL4Rα or IL13Rα1 in SNU308 cells. ( A ) Cell viability (WST-1 assay) was measured for 24, 48, and 72 h after transfection of siRNA. The percentage of SNU308 cell viability transfected with siRNAs against IL4Rα and IL13Rα1 was compared with the cells transfected with control siRNA (cell viability = 100%) at 24 h after transfection. ( B ) The number of viable cells was counted with hemocytometer for 24, 48, and 72 h after transfection of siRNA. ( C ) After transfection of siRNA, cells were grown for 14 days in fresh media containing 10% FBS. Crystal violet staining was performed to visualize colonies. ( D ) Cell morphological image was captured by microscopy for 24, 48, and 72 h after transfection of siRNA. * p < 0.05; asterisks mean a significant difference between control siRNA group and IL4Rα or IL13Rα1 siRNA group. Results are mean ± SD of at least three independent experiments.

Article Snippet: 5 uL of control, IL4Rα, or IL13Rα1 siRNA (Santa Cruz, the concentration of siRNA stock solution is 10 uM) and 15 µL of lipofectamine 2000 (Waltham, MA, USA) were added to media respectively and incubated for 15 min. After incubation, the media containing siRNA and the media containing lipofectamine 2000 were mixed and incubated for 15 min. After 15 min of incubation, cells were washed twice with PBS and the siRNA and lipofectamine 2000 mixtures were added to cells.

Techniques: WST-1 Assay, Transfection, Staining, Microscopy

Journal: Journal of Personalized Medicine

Article Title: IL4Rα and IL13Rα1 Are Involved in the Development of Human Gallbladder Cancer

doi: 10.3390/jpm12020249

Figure Lengend Snippet: Cell cycle analysis and measurement of apoptosis by knockdown of IL4Rα or IL13Rα1 in SNU308 cells. ( A ) Cell cycle population was determined by Muse Cell Analyzer and Muse Cell Cycle Kit for 48 h after transfection of siRNA. ( B ) Annexin V staining was analyzed by Muse Cell Analyzer and Muse Annexin V and Dead Cell Kit for 48 h after transfection of siRNA. ( C ) Caspase-3 and -7 activities were measured by Muse Cell Analyzer and Muse Caspase-3/7 Kit for 48 h after transfection of siRNA. ( D ) Depolarization of mitochondria was detected by Muse Cell Analyzer and Muse Mito-Potential Kit for 48 h after transfection of siRNA. Quantitative graph was added to each result. * p < 0.05; asterisks mean a significant difference between control siRNA group and IL4Rα or IL13Rα1 siRNA group. Results are mean ± SD of at least three independent experiments.

Article Snippet: 5 uL of control, IL4Rα, or IL13Rα1 siRNA (Santa Cruz, the concentration of siRNA stock solution is 10 uM) and 15 µL of lipofectamine 2000 (Waltham, MA, USA) were added to media respectively and incubated for 15 min. After incubation, the media containing siRNA and the media containing lipofectamine 2000 were mixed and incubated for 15 min. After 15 min of incubation, cells were washed twice with PBS and the siRNA and lipofectamine 2000 mixtures were added to cells.

Techniques: Cell Cycle Assay, Transfection, Staining

Journal: Journal of Personalized Medicine

Article Title: IL4Rα and IL13Rα1 Are Involved in the Development of Human Gallbladder Cancer

doi: 10.3390/jpm12020249

Figure Lengend Snippet: Visualization of DNA damage by knockdown of IL4Rα or IL13Rα1 in SNU308 cells. ( A ) Comet assay was conducted to visualize DNA damage in nucleus for 48 h after transfection of siRNA. ( B ) Blue color represents nucleus stained by Hoechst and green color represent TUNEL-positive nucleus for 48 h after transfection of siRNA. DNA damage of cell was observed by fluorescence microscopy. Green color in the cytogram means live cells. Quantitative graph was added to each result. * p < 0.05; asterisks mean a significant difference between control siRNA group and IL4Rα or IL13Rα1 siRNA group. Results are mean ± SD of at least three independent experiments.

Article Snippet: 5 uL of control, IL4Rα, or IL13Rα1 siRNA (Santa Cruz, the concentration of siRNA stock solution is 10 uM) and 15 µL of lipofectamine 2000 (Waltham, MA, USA) were added to media respectively and incubated for 15 min. After incubation, the media containing siRNA and the media containing lipofectamine 2000 were mixed and incubated for 15 min. After 15 min of incubation, cells were washed twice with PBS and the siRNA and lipofectamine 2000 mixtures were added to cells.

Techniques: Single Cell Gel Electrophoresis, Transfection, Staining, TUNEL Assay, Fluorescence, Microscopy

Journal: Journal of Personalized Medicine

Article Title: IL4Rα and IL13Rα1 Are Involved in the Development of Human Gallbladder Cancer

doi: 10.3390/jpm12020249

Figure Lengend Snippet: Western blotting analysis of proteins in SNU308 cells transfected with IL4Rα or IL13Rα1. ( A ) Total cell lysates Western blotting analysis in SNU308 for 48 h after transfection of siRNA. β-actin was used for a gel-loading control. ( B ) Nuclear fractional Western blotting analysis in SNU308 for 48 h after transfection of siRNA. GAPDH and Lamin A/C were used for a gel loading control for the cytosol and nuclear protein fractions, respectively. These results were representative images from three independent experiments.

Article Snippet: 5 uL of control, IL4Rα, or IL13Rα1 siRNA (Santa Cruz, the concentration of siRNA stock solution is 10 uM) and 15 µL of lipofectamine 2000 (Waltham, MA, USA) were added to media respectively and incubated for 15 min. After incubation, the media containing siRNA and the media containing lipofectamine 2000 were mixed and incubated for 15 min. After 15 min of incubation, cells were washed twice with PBS and the siRNA and lipofectamine 2000 mixtures were added to cells.

Techniques: Western Blot, Transfection

Journal: Journal of Personalized Medicine

Article Title: IL4Rα and IL13Rα1 Are Involved in the Development of Human Gallbladder Cancer

doi: 10.3390/jpm12020249

Figure Lengend Snippet: Anti-proliferative effect by AZD1480 treatment in SNU308 cells. ( A ) Cell viability (WST-1 assay) was measured for 24, 48, and 72 h after AZD1480 treatment. The percentage of SNU308 cell viability transfected with siRNAs against IL4Rα and IL13Rα1 was compared with the cells transfected with control siRNA (cell viability = 100%) at 24 h after transfection. ( B ) The number of viable cells was counted with hemocytometer for 24, 48, and 72 h after AZD1480 treatment. ( C ) After AZD1480 treatment, cells were grown for 14 days in fresh media containing 10% FBS. Crystal violet staining was performed to visualize colonies. ( D ) Cell morphological image was captured by microscopy for 24, 48, and 72 h after AZD1480 treatment. * p < 0.05; asterisks mean a significant difference between control group and treatment group. Results are mean ± SD of at least three independent experiments.

Article Snippet: 5 uL of control, IL4Rα, or IL13Rα1 siRNA (Santa Cruz, the concentration of siRNA stock solution is 10 uM) and 15 µL of lipofectamine 2000 (Waltham, MA, USA) were added to media respectively and incubated for 15 min. After incubation, the media containing siRNA and the media containing lipofectamine 2000 were mixed and incubated for 15 min. After 15 min of incubation, cells were washed twice with PBS and the siRNA and lipofectamine 2000 mixtures were added to cells.

Techniques: WST-1 Assay, Transfection, Staining, Microscopy

Journal: Science Advances

Article Title: Attenuation of Dupuytren’s fibrosis via targeting of the STAT1 modulated IL-13Rα1 response

doi: 10.1126/sciadv.aaz8272

Figure Lengend Snippet: Dupuytren’s myofibroblasts were pretreated with anti–IL13Rα1 before exposure to IL-13 (100 ng/ml). ( A ) Phosphorylation of STAT6. ( B ) Proliferation of Dupuytren’s myofibroblasts. ( C ) Effect on collagen 1 production and tenascin-C, periostin, and IL-13Rα2 gene expression. mRNA gene expression expressed as fold change following normalization to housekeeping gene (GAPDH) and then to relevant untreated cells. All results are means ± SEM, n = 6; * indicates significant difference from untreated cells, * P < 0.05, ** P < 0.01, *** P < 0.001. # P < 0.05, ## P < 0.01, ### P < 0.001. ( D ) IL-13Rα1 protein and transcript expression in control fibroblasts and Dupuytren’s myofibroblasts. Representative flow cytometry histogram of control fibroblasts (blue line) and Dupuytren’s myofibroblasts (red line). Graphs illustrates MFI or 2 −ΔCT (relative to GAPDH) of IL-13Rα1, means ± SEM, n = 6; ** P < 0.01.

Article Snippet: Using the publicly available UCSC (University of California Santa Cruz) genome browser database, we identified a number of potential binding sites of interest for STAT1 and SPI1 [a key regulator of myofibroblast differentiation in fibrosis ( )] on the IL-13Rα1 gene locus ( ).

Techniques: Expressing, Flow Cytometry

Journal: Science Advances

Article Title: Attenuation of Dupuytren’s fibrosis via targeting of the STAT1 modulated IL-13Rα1 response

doi: 10.1126/sciadv.aaz8272

Figure Lengend Snippet: ( A ) Control fibroblasts exposed to IFN-γ and/or TGF-β. IL-13Rα1 transcript following treatment. Representative flow cytometry histogram of IL-13Rα1 expression on the surface of untreated, and graph illustrates MFI of IL-13Rα1 after treatment. All results are means ± SEM; mRNA gene expression expressed as fold change following normalization to housekeeping gene (GAPDH) and then to relevant untreated cells, n = 6; * indicates significant difference from untreated cells, * P < 0.05. # P < 0.05. ( B ) Identification of sites of interest on IL-13Rα1 gene locus: nonspecific (blue), enhancer (green), promoter (red), and intronic site (purple). ( C ) Quantitative polymerase chain reaction of ChIP representing percent enriched binding of SPI1 and STAT1 at different sites of interest of interest on the IL-13Rα1 gene. Graph demonstrates the percent enriched following subtraction of nonspecific enrichment, means ± SEM, n = 4; * P < 0.05.

Article Snippet: Using the publicly available UCSC (University of California Santa Cruz) genome browser database, we identified a number of potential binding sites of interest for STAT1 and SPI1 [a key regulator of myofibroblast differentiation in fibrosis ( )] on the IL-13Rα1 gene locus ( ).

Techniques: Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction, Binding Assay

Journal: Science Advances

Article Title: Attenuation of Dupuytren’s fibrosis via targeting of the STAT1 modulated IL-13Rα1 response

doi: 10.1126/sciadv.aaz8272

Figure Lengend Snippet: ( A ) STAT1 phosphorylation and IL-13 secretion from human mast cells following IFN-γ and TGF-β exposure. Cells were pretreated with tofacitinib or vehicle control [0.001% dimethyl sulfoxide (DMSO)] for 30 min. ( B ) STAT1 phosphorylation and IL-13Rα1 transcript and protein expression following IFN-γ and TGF-β treatment. Control fibroblasts were pretreated with tofacitinib or vehicle control (0.001% DMSO) for 30 min before cytokine stimulation. ( C ) Dupuytren’s myofibroblasts were pretreated with tofacitinib or vehicle control (0.001% DMSO) and then exposed to IL-13 (100 ng/ml). STAT6 phosphorylation, myofibroblast proliferation, and periostin, tenascin-C, and IL-13Rα2 transcript levels. All results are means ± SEM; mRNA gene expression expressed as fold change following normalization to housekeeping gene (GAPDH) and then to relevant untreated cells, n > 4; * indicates significant difference from untreated cells, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.001. # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001.

Article Snippet: Using the publicly available UCSC (University of California Santa Cruz) genome browser database, we identified a number of potential binding sites of interest for STAT1 and SPI1 [a key regulator of myofibroblast differentiation in fibrosis ( )] on the IL-13Rα1 gene locus ( ).

Techniques: Expressing

Journal: Cancers

Article Title: Interleukin4Rα (IL4Rα) and IL13Rα1 Are Associated with the Progress of Renal Cell Carcinoma through Janus Kinase 2 (JAK2)/Forkhead Box O3 (FOXO3) Pathways

doi: 10.3390/cancers11091394

Figure Lengend Snippet: Immunohistochemical expression and survival analysis for the expression of interleukin4 receptor α (IL4Rα) and interleukin13 receptor α1 (IL13Rα1) in clear cell renal cell carcinoma (CCRCC). ( A ) Immunohistochemical expression of IL4Rα and IL13Rα1 in CCRCC tissue. Original magnification, ×400. ( B ) Analysis of sensitivity and specificity of the immunohistochemical staining score of IL4Rα and IL13Rα1 in CCRCC for the event of cancer-specific survival (death of the patient by CCRCC) by receiver operator characteristic curves. The cutoff points were determined at the highest area under the curve (AUC). Red arrow indicates a cutoff point for the IL4Rα immunostaining and blue arrowhead indicates a cut-off point for the IL13Rα1 immunostaining. ( C ) Kaplan-Meier survival analysis for cancer-specific survival and relapse-free survival according to the immunohistochemical positivity for IL4Rα and IL13Rα1 in 199 CCRCC. ( D ) Kaplan-Meier survival analysis in IL4Rα−/IL13Rα1−, IL4Rα−/IL13Rα1+, IL4Rα+/IL13Rα1−, and IL4Rα+/IL13Rα1+ subgroups according to the positivity for IL4Rα and IL13Rα1 expressions. ( E ) Kaplan-Meier survival analysis in two subgroups according to the co-expression patterns of IL4Rα and IL13Rα1; favorable (IL4Rα−/IL13Rα1−, IL4Rα−/IL13Rα1+, or IL4Rα+/IL13Rα1−) and poor prognostic (IL4Rα+/IL13Rα1+) subgroups. 5y-cancer-specific survival (CSS); 5-year cancer-specific survival rate, 10y-CSS; 10-year cancer-specific survival rate; 5y-RFS; five-year relapse-free survival rate, 10y- relapse-free survival (RFS); 10-year relapse-free survival rate.

Article Snippet: After incubation, cells were transfected with siRNAs (control (Cat. No.: sc37007), IL4Rα (Cat. No.: sc-35661), IL13Rα1 (Cat. No.: sc-63337), or JAK2 (Cat. No.: sc-39099), 1 nM (Santa Cruz Biotechnology)) or plasmid DNAs (pECE empty/Flag-FOXO3 or pCMV3-C-HA empty/HA-JAK2 plasmid DNA, 1 µg) to 3 µL of Lipofectamine 2000 (Invitrogen) in 300 µL of serum-free media for 6 h at 37 °C in a humidified incubator containing 5% CO 2 in air.

Techniques: Immunohistochemical staining, Expressing, Staining, Immunostaining

Journal: Cancers

Article Title: Interleukin4Rα (IL4Rα) and IL13Rα1 Are Associated with the Progress of Renal Cell Carcinoma through Janus Kinase 2 (JAK2)/Forkhead Box O3 (FOXO3) Pathways

doi: 10.3390/cancers11091394

Figure Lengend Snippet: Clinicopathologic variables and the expression of interleukin4 receptor α (IL4Rα) and interleukin13 receptor α1 (IL13Rα1) in clear cell renal cell carcinoma (CCRCC) patients.

Article Snippet: After incubation, cells were transfected with siRNAs (control (Cat. No.: sc37007), IL4Rα (Cat. No.: sc-35661), IL13Rα1 (Cat. No.: sc-63337), or JAK2 (Cat. No.: sc-39099), 1 nM (Santa Cruz Biotechnology)) or plasmid DNAs (pECE empty/Flag-FOXO3 or pCMV3-C-HA empty/HA-JAK2 plasmid DNA, 1 µg) to 3 µL of Lipofectamine 2000 (Invitrogen) in 300 µL of serum-free media for 6 h at 37 °C in a humidified incubator containing 5% CO 2 in air.

Techniques: Expressing

Journal: Cancers

Article Title: Interleukin4Rα (IL4Rα) and IL13Rα1 Are Associated with the Progress of Renal Cell Carcinoma through Janus Kinase 2 (JAK2)/Forkhead Box O3 (FOXO3) Pathways

doi: 10.3390/cancers11091394

Figure Lengend Snippet: Univariate Cox regression analysis of cancer-specific survival (CSS) and relapse-free survival (RFS) in clear cell renal cell carcinoma (CCRCC) patients.

Article Snippet: After incubation, cells were transfected with siRNAs (control (Cat. No.: sc37007), IL4Rα (Cat. No.: sc-35661), IL13Rα1 (Cat. No.: sc-63337), or JAK2 (Cat. No.: sc-39099), 1 nM (Santa Cruz Biotechnology)) or plasmid DNAs (pECE empty/Flag-FOXO3 or pCMV3-C-HA empty/HA-JAK2 plasmid DNA, 1 µg) to 3 µL of Lipofectamine 2000 (Invitrogen) in 300 µL of serum-free media for 6 h at 37 °C in a humidified incubator containing 5% CO 2 in air.

Techniques:

Journal: Cancers

Article Title: Interleukin4Rα (IL4Rα) and IL13Rα1 Are Associated with the Progress of Renal Cell Carcinoma through Janus Kinase 2 (JAK2)/Forkhead Box O3 (FOXO3) Pathways

doi: 10.3390/cancers11091394

Figure Lengend Snippet: Multivariate Cox regression analysis of CSS and RFS in CCRCC patients.

Article Snippet: After incubation, cells were transfected with siRNAs (control (Cat. No.: sc37007), IL4Rα (Cat. No.: sc-35661), IL13Rα1 (Cat. No.: sc-63337), or JAK2 (Cat. No.: sc-39099), 1 nM (Santa Cruz Biotechnology)) or plasmid DNAs (pECE empty/Flag-FOXO3 or pCMV3-C-HA empty/HA-JAK2 plasmid DNA, 1 µg) to 3 µL of Lipofectamine 2000 (Invitrogen) in 300 µL of serum-free media for 6 h at 37 °C in a humidified incubator containing 5% CO 2 in air.

Techniques:

Journal: Cancers

Article Title: Interleukin4Rα (IL4Rα) and IL13Rα1 Are Associated with the Progress of Renal Cell Carcinoma through Janus Kinase 2 (JAK2)/Forkhead Box O3 (FOXO3) Pathways

doi: 10.3390/cancers11091394

Figure Lengend Snippet: Anti-cancer effect by transfection of siRNA against IL4Rα or IL13Rα1 in A498 and ACHN cells. Time-dependent anti-cancer effect by transfection of siRNA against IL4Rα or IL13Rα1 in A498 and ACHN cells for 24, 48, and 72 h incubation after transfection. Cell viability and proliferation rate was determined by WST-1 assay ( A ) and cell counting assay ( B ), respectively. This result is representative data from three biological replicates, and the error bar indicates standard error (STE). * indicates the p -value < 0.05. ( C ) Anti-colony formation ability by transfection of siRNA against IL4Rα or IL13Rα1 in A498 and ACHN cells was determined by colony formation assay for 14 days after transfection. These results are representative data from three biological replicates. Apoptosis in A498 and ACHN cells transfected with siRNA against IL4Rα or IL13Rα1 for 48 h after transfection was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay ( D ) and Annexin V staining analysis ( E ). Cell cycle arrest was determined by cell cycle analysis ( F ). This result is representative data from three biological replicates. ( G ) Western blotting analysis of proteins related to apoptosis and cell cycle arrest in A498 and ACHN cells transfected with siRNA against IL4Rα or IL13Rα1 for 48 h after transfection. β-actin was used for a gel-loading control. Magnification for ( D ): ×20.

Article Snippet: After incubation, cells were transfected with siRNAs (control (Cat. No.: sc37007), IL4Rα (Cat. No.: sc-35661), IL13Rα1 (Cat. No.: sc-63337), or JAK2 (Cat. No.: sc-39099), 1 nM (Santa Cruz Biotechnology)) or plasmid DNAs (pECE empty/Flag-FOXO3 or pCMV3-C-HA empty/HA-JAK2 plasmid DNA, 1 µg) to 3 µL of Lipofectamine 2000 (Invitrogen) in 300 µL of serum-free media for 6 h at 37 °C in a humidified incubator containing 5% CO 2 in air.

Techniques: Transfection, Incubation, WST-1 Assay, Cell Counting, Colony Assay, TUNEL Assay, Staining, Cell Cycle Assay, Western Blot

Journal: Cancers

Article Title: Interleukin4Rα (IL4Rα) and IL13Rα1 Are Associated with the Progress of Renal Cell Carcinoma through Janus Kinase 2 (JAK2)/Forkhead Box O3 (FOXO3) Pathways

doi: 10.3390/cancers11091394

Figure Lengend Snippet: Interaction between Janus kinase 2 (JAK2) and Forkhead box O3 (FOXO3) protein in A498 and ACHN. ( A ) Silencing IL4Rα in A498 and ACHN cells reduced the interaction between JAK2 and FOXO3. Cells were transfected with control siRNA or siRNA against IL4Rα, and then cell lysates were immunoprecipitated with the anti-pJAK2 antibody. The immunoprecipitated proteins were resolved on the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted by anti-IL4Rα, IL13Rα1, FOXO3, and pJAK2 antibody. The light chain of IgG was used for the loading control. ( B ) 293T cells were co-transfected with HA- JAK2 and Flag-FOXO3 (O.E.) or a control plasmid DNA (pCMV3-C-HA and pECE, Con.) as indicated. Then cell lysates were immunoprecipitated with anti-Flag, FOXO3, HA, JAK2, or pTyr antibody. The immunoprecipitated proteins were resolved on the SDS-PAGE and immunoblotted by the indicated antibody, respectively. The light chain of IgG and Coomassie Blue staining of SDS-PAGE were used for the loading control. ( C ) Silencing JAK2 in A498 and ACHN cells increased the expression of FOXO3. Cells were transfected with control siRNA or siRNA against JAK2, and then cell lysates were resolved on the SDS-PAGE and immunoblotted by anti-FOXO3 and JAK2 antibody. β-actin was used for the loading control. ( D ) Pharmacological inhibition of JAK2 in A498 and ACHN cells increased the expression of FOXO3. Cells were treated with dimethyl sulfoxide (DMSO) vehicle control or the indicated concentration of AZD1480, and then cell lysates were resolved on the SDS-PAGE and immunoblotted by anti-FOXO3, pJAK2, and JAK2 antibody. β-actin was used for the loading control.

Article Snippet: After incubation, cells were transfected with siRNAs (control (Cat. No.: sc37007), IL4Rα (Cat. No.: sc-35661), IL13Rα1 (Cat. No.: sc-63337), or JAK2 (Cat. No.: sc-39099), 1 nM (Santa Cruz Biotechnology)) or plasmid DNAs (pECE empty/Flag-FOXO3 or pCMV3-C-HA empty/HA-JAK2 plasmid DNA, 1 µg) to 3 µL of Lipofectamine 2000 (Invitrogen) in 300 µL of serum-free media for 6 h at 37 °C in a humidified incubator containing 5% CO 2 in air.

Techniques: Transfection, Immunoprecipitation, Polyacrylamide Gel Electrophoresis, SDS Page, Plasmid Preparation, Staining, Expressing, Inhibition, Concentration Assay

Journal: Cancers

Article Title: Interleukin4Rα (IL4Rα) and IL13Rα1 Are Associated with the Progress of Renal Cell Carcinoma through Janus Kinase 2 (JAK2)/Forkhead Box O3 (FOXO3) Pathways

doi: 10.3390/cancers11091394

Figure Lengend Snippet: A diagram describes the role of IL4R complex (a heterodimeric complex of IL4Rα and IL13Rα1) in promoting RCC tumorigenesis via phosphorylation of tyrosine residue in FOXO3 by activation of JAK2 leading to loss of the tumor-suppressive transcriptional activity.

Article Snippet: After incubation, cells were transfected with siRNAs (control (Cat. No.: sc37007), IL4Rα (Cat. No.: sc-35661), IL13Rα1 (Cat. No.: sc-63337), or JAK2 (Cat. No.: sc-39099), 1 nM (Santa Cruz Biotechnology)) or plasmid DNAs (pECE empty/Flag-FOXO3 or pCMV3-C-HA empty/HA-JAK2 plasmid DNA, 1 µg) to 3 µL of Lipofectamine 2000 (Invitrogen) in 300 µL of serum-free media for 6 h at 37 °C in a humidified incubator containing 5% CO 2 in air.

Techniques: Activation Assay, Activity Assay

Journal: The Journal of Experimental Medicine

Article Title: A role for sex chromosome complement in the female bias in autoimmune disease

doi: 10.1084/jem.20070850

Figure Lengend Snippet: Effect of sex chromosome complement on IL-13Rα2 expression. Freshly isolated splenocytes from PLP 139–151 immunized SJL mice of the XX or the XY − sex chromosome complement were analyzed for the expression of IL-13Rα2 (top) and IL-13Rα1 (bottom) on B cells (CD19 + ), macrophages (CD11b + ), dendritic cells (CD11c + ), and myeloid dendritic cells (CD11c + CD11b + ) by flow cytometry. XX, thick red lines; XY − , blue lines; shaded area, negative control. The mean fluorescent intensities are indicated on FACS plots (red, XX; blue, XY − ). IL-13Rα2 expression was significantly higher on macrophages, dendritic cells, and myeloid dendritic cells in XX mice than in XY − mice. *, P < 0.05; n = 3 each. Results represent two independent experiments.

Article Snippet: IL-13Rα1 expression was detected using FITC-conjugated antibody H300 (Santa Cruz Biotechnology).

Techniques: Expressing, Isolation, Flow Cytometry, Negative Control