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recombinant murine wisp1 (R&D Systems)

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R&D Systems recombinant murine wisp1

(A) <t>Wisp1</t> mRNA expression in primary adult mouse cardiac fibroblasts following TGFβ1 (10 ng/mL) treatment for 24–72 h. (B) Representative immunofluorescence images of α-SMA (green), vimentin (red), and DAPI (blue) after 72 h treatment with vehicle (Ctrl), WISP1 (500 ng/mL), TGFβ1 (10 ng/mL), or WISP1 + TGFβ1. (C, D) Representative Western blot (C) and densitometric quantification (D) of α-SMA normalized to GAPDH. Data are mean ± SEM from ≥3 independent isolations. Statistical analysis by one-way ANOVA with Tukey’s post hoc test; *p < 0.05. Data are mean ± SEM from ≥3 independent isolations. Statistical analysis by one-way ANOVA with Tukey’s post hoc test; *p < 0.05.

Recombinant Murine Wisp1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 96/100, based on 169 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/recombinant murine wisp1/product/R&D Systems
Average 96 stars, based on 169 article reviews

recombinant murine wisp1 - by Bioz Stars, 2026-05

96/100 stars

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1) Product Images from "WISP1 drives a mechanically active immune modulatory and proliferative cardiac myofibroblast state"

Article Title: WISP1 drives a mechanically active immune modulatory and proliferative cardiac myofibroblast state

Journal: bioRxiv

doi: 10.64898/2026.02.17.706476

(A) Wisp1 mRNA expression in primary adult mouse cardiac fibroblasts following TGFβ1 (10 ng/mL) treatment for 24–72 h. (B) Representative immunofluorescence images of α-SMA (green), vimentin (red), and DAPI (blue) after 72 h treatment with vehicle (Ctrl), WISP1 (500 ng/mL), TGFβ1 (10 ng/mL), or WISP1 + TGFβ1. (C, D) Representative Western blot (C) and densitometric quantification (D) of α-SMA normalized to GAPDH. Data are mean ± SEM from ≥3 independent isolations. Statistical analysis by one-way ANOVA with Tukey’s post hoc test; *p < 0.05. Data are mean ± SEM from ≥3 independent isolations. Statistical analysis by one-way ANOVA with Tukey’s post hoc test; *p < 0.05.

Figure Legend Snippet: (A) Wisp1 mRNA expression in primary adult mouse cardiac fibroblasts following TGFβ1 (10 ng/mL) treatment for 24–72 h. (B) Representative immunofluorescence images of α-SMA (green), vimentin (red), and DAPI (blue) after 72 h treatment with vehicle (Ctrl), WISP1 (500 ng/mL), TGFβ1 (10 ng/mL), or WISP1 + TGFβ1. (C, D) Representative Western blot (C) and densitometric quantification (D) of α-SMA normalized to GAPDH. Data are mean ± SEM from ≥3 independent isolations. Statistical analysis by one-way ANOVA with Tukey’s post hoc test; *p < 0.05. Data are mean ± SEM from ≥3 independent isolations. Statistical analysis by one-way ANOVA with Tukey’s post hoc test; *p < 0.05.

Techniques Used: Expressing, Immunofluorescence, Western Blot

Primary adult mouse cardiac fibroblasts were embedded in floating collagen gels and treated for 24 h with vehicle (Ctrl), WISP1 (500 ng/mL), TGFβ1 (10 ng/mL), or WISP1 + TGFβ1. (A) Representative gel images at 0 h and 24 h. (B) Quantification of gel area (% contraction; n = 4 isolations, 2 males & 2 females). (C–D) Confluent fibroblast monolayers were scratched and imaged every 2 hours for 24 hours using the same treatment conditions. (C) Representative bright-field images at time point 0 and 14. (D) Quantification of wound closure (% of gap closed relative to time 0; n = 8 isolations, 4 male & 4 female). Data are mean ± SEM. One-way ANOVA with Tukey’s multiple comparisons; *p ≤ 0.05.

Figure Legend Snippet: Primary adult mouse cardiac fibroblasts were embedded in floating collagen gels and treated for 24 h with vehicle (Ctrl), WISP1 (500 ng/mL), TGFβ1 (10 ng/mL), or WISP1 + TGFβ1. (A) Representative gel images at 0 h and 24 h. (B) Quantification of gel area (% contraction; n = 4 isolations, 2 males & 2 females). (C–D) Confluent fibroblast monolayers were scratched and imaged every 2 hours for 24 hours using the same treatment conditions. (C) Representative bright-field images at time point 0 and 14. (D) Quantification of wound closure (% of gap closed relative to time 0; n = 8 isolations, 4 male & 4 female). Data are mean ± SEM. One-way ANOVA with Tukey’s multiple comparisons; *p ≤ 0.05.

Techniques Used:

Image Search Results

Differential expression of WISP1 in normal and tumor tissue samples. (A) Expression of WISP1 in pan-cancer tissues and adjacent normal tissues via TIMER2.0. (B) Expression levels of WISP1 in ESCA from GEPIA2 (|Log2FC|> 1, P < 0.01, log scale: log2 (TPM + 1), Jitter Size: 0.4; T: Tumor, N: Normal). (C) Expression of WISP1 mRNA in the ESCC dataset. (D–G) Results of IHC and WB analyses using 12 paired ESCC tissues and adjacent control samples (T: Tumor, N: Normal; scale bars = 100μm). (H) ROC curves predicting the prognostic ability of high WISP1 expression for 1-year, 3-year, and 5-year patient survival. *p < 0.05; **p < 0.01; ***p < 0.001,;****P < 0.0001.

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: Differential expression of WISP1 in normal and tumor tissue samples. (A) Expression of WISP1 in pan-cancer tissues and adjacent normal tissues via TIMER2.0. (B) Expression levels of WISP1 in ESCA from GEPIA2 (|Log2FC|> 1, P < 0.01, log scale: log2 (TPM + 1), Jitter Size: 0.4; T: Tumor, N: Normal). (C) Expression of WISP1 mRNA in the ESCC dataset. (D–G) Results of IHC and WB analyses using 12 paired ESCC tissues and adjacent control samples (T: Tumor, N: Normal; scale bars = 100μm). (H) ROC curves predicting the prognostic ability of high WISP1 expression for 1-year, 3-year, and 5-year patient survival. *p < 0.05; **p < 0.01; ***p < 0.001,;****P < 0.0001.

Article Snippet: Primary fibroblasts were infected with shWISP1 lentivirus or treated with recombinant human WISP1 (rhWISP1) protein( NP_003873.1 , Sino Biological, China; endotoxin level <0.1 EU/μg, as determined by Limulus Amebocyte Lysate assay), followed by three washes with PBS.

Techniques: Quantitative Proteomics, Expressing, Control

The impact of WISP1 expression on the survival of ESCC patients. (A–C) Kaplan-Meier analysis of overall survival for high-expression versus low-expression groups in the datasets GSE53624 , GSE53625 , and TCGA. (D–F) Time-dependent ROC analysis for patients in the datasets.

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: The impact of WISP1 expression on the survival of ESCC patients. (A–C) Kaplan-Meier analysis of overall survival for high-expression versus low-expression groups in the datasets GSE53624 , GSE53625 , and TCGA. (D–F) Time-dependent ROC analysis for patients in the datasets.

Techniques: Expressing

Identification of DEGs in ESCC, functional enrichment analysis, and functional annotation of WISP1. (A) Volcano plot shows the DEGs from four datasets. (B) A Venn diagram shows genes that are differentially expressed across all four datasets. (C) GO and KEGG analyses reveal the potential biological mechanisms of DEGs in the four datasets. (D, E) GeneMANIA and STRING databases identify target proteins and genes associated with WISP1, followed by enrichment analysis of the functions of these proteins and genes. (F) Visualization of enrichment analysis results.

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: Identification of DEGs in ESCC, functional enrichment analysis, and functional annotation of WISP1. (A) Volcano plot shows the DEGs from four datasets. (B) A Venn diagram shows genes that are differentially expressed across all four datasets. (C) GO and KEGG analyses reveal the potential biological mechanisms of DEGs in the four datasets. (D, E) GeneMANIA and STRING databases identify target proteins and genes associated with WISP1, followed by enrichment analysis of the functions of these proteins and genes. (F) Visualization of enrichment analysis results.

Techniques: Functional Assay

Tumor microenvironment and immune cell infiltration analysis in the GSE53624 dataset. (A) Differences in the abundance of infiltrating immune cells between high-expression and low-expression groups. (B–D) Differences in stromal scores, immune scores, and estimate scores between high-expression and low-expression groups. (E) Evaluation of the expression of immune checkpoint molecules (CD274, PDCD1, TIGIT, CD276, CTLA4, LAG3) between high-risk and low-risk groups. (F) Correlation between WISP1 expression and immune checkpoint molecules (CD274, PDCD1, TIGIT, CD276, CTLA4, LAG3). Blue indicates positive correlation, red indicates negative correlation, and the numbers inside the boxes represent the magnitude of the correlation. *p < 0.05; **p < 0.01; ***p < 0.001; ****P < 0.0001.

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: Tumor microenvironment and immune cell infiltration analysis in the GSE53624 dataset. (A) Differences in the abundance of infiltrating immune cells between high-expression and low-expression groups. (B–D) Differences in stromal scores, immune scores, and estimate scores between high-expression and low-expression groups. (E) Evaluation of the expression of immune checkpoint molecules (CD274, PDCD1, TIGIT, CD276, CTLA4, LAG3) between high-risk and low-risk groups. (F) Correlation between WISP1 expression and immune checkpoint molecules (CD274, PDCD1, TIGIT, CD276, CTLA4, LAG3). Blue indicates positive correlation, red indicates negative correlation, and the numbers inside the boxes represent the magnitude of the correlation. *p < 0.05; **p < 0.01; ***p < 0.001; ****P < 0.0001.

Techniques: Expressing

Gene set enrichment analysis (GSEA) of hallmark pathways for esophageal squamous cell carcinoma (ESCC) patients stratified by WISP1 expression levels. (A–C) Enrichment patterns of associated pathways in the GSE53624 , GSE53625 , and TCGA datasets. The analyses were evaluated using a significance threshold corrected for the False Discovery Rate (FDR).

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: Gene set enrichment analysis (GSEA) of hallmark pathways for esophageal squamous cell carcinoma (ESCC) patients stratified by WISP1 expression levels. (A–C) Enrichment patterns of associated pathways in the GSE53624 , GSE53625 , and TCGA datasets. The analyses were evaluated using a significance threshold corrected for the False Discovery Rate (FDR).

Techniques: Expressing

Computational prediction of drug sensitivity for ESCC patients stratified by WISP1 expression levels. Using WISP1 expression grouping derived from the GSE53624 dataset, the “oncoPredict” R package was employed to evaluate the drug response spectrum. **** indicates statistical significance at p < 0.0001.

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: Computational prediction of drug sensitivity for ESCC patients stratified by WISP1 expression levels. Using WISP1 expression grouping derived from the GSE53624 dataset, the “oncoPredict” R package was employed to evaluate the drug response spectrum. **** indicates statistical significance at p < 0.0001.

Techniques: Expressing, Derivative Assay

Functional characterization of WISP1 in a subpopulation of fibroblasts. (A) Gene Set Enrichment Analysis (GSEA) shows enrichment results of differentially expressed genes between WISP1_Fib_Negative and WISP1_Fib_Positive clusters within the c5.all.v2024.1.Hs.symbols gene set; (B, C) display the enrichment results of DEGs in the h.all.v7.1.symbols gene set. (D) Proportional distribution of WISP1_Fib_Negative and WISP1_Fib_Positive clusters in normal esophageal tissues and ESCC samples. (E, F) Comparative analysis of intercellular communication signal intensity among fibroblast subpopulations. (G, H) Molecular stratification of fibroblasts using lineage-specific markers (DCN/Decorin, IGFBP6/Insulin Like Growth Factor Binding Protein 6, MFAP5/Microfibril Associated Protein 5, ACTA2/Actin Alpha 2 Smooth Muscle, TAGLN/Transgelin, CTHRC1/Collagen Triple Helix Repeat Containing 1)into NFs and CAFs subtypes. (I) Validation of CAF-specific markers (FAP, COL1A1, COL3A1, COL4A1, COL10A1, MMP1, MMP11, MMP14) and expression patterns of WISP1 in CAFs via single-cell RNA sequencing.

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: Functional characterization of WISP1 in a subpopulation of fibroblasts. (A) Gene Set Enrichment Analysis (GSEA) shows enrichment results of differentially expressed genes between WISP1_Fib_Negative and WISP1_Fib_Positive clusters within the c5.all.v2024.1.Hs.symbols gene set; (B, C) display the enrichment results of DEGs in the h.all.v7.1.symbols gene set. (D) Proportional distribution of WISP1_Fib_Negative and WISP1_Fib_Positive clusters in normal esophageal tissues and ESCC samples. (E, F) Comparative analysis of intercellular communication signal intensity among fibroblast subpopulations. (G, H) Molecular stratification of fibroblasts using lineage-specific markers (DCN/Decorin, IGFBP6/Insulin Like Growth Factor Binding Protein 6, MFAP5/Microfibril Associated Protein 5, ACTA2/Actin Alpha 2 Smooth Muscle, TAGLN/Transgelin, CTHRC1/Collagen Triple Helix Repeat Containing 1)into NFs and CAFs subtypes. (I) Validation of CAF-specific markers (FAP, COL1A1, COL3A1, COL4A1, COL10A1, MMP1, MMP11, MMP14) and expression patterns of WISP1 in CAFs via single-cell RNA sequencing.

Techniques: Functional Assay, Binding Assay, Biomarker Discovery, Expressing, RNA Sequencing

WISP1 regulates cancer-associated fibroblast function and extracellular matrix remodeling. (A, B) Immunofluorescence multiplex staining of α-smooth muscle actin (αSMA) and fibroblast activation protein (FAP) in paired cancer-associated fibroblasts (CAFs, tumor-derived) and normal fibroblasts (NFs, adjacent non-tumor tissues) (scale bars = 50μm). (C) Western blot (WB) analysis of WISP1, αSMA, and FAP expression in CAFs versus NFs. (D, E) Lentiviral short hairpin RNA (shRNA)-mediated WISP1 knockdown in CAFs, validated by quantitative reverse transcription PCR (qRT-PCR) and WB. (F–K) Functional characterization of CAF proliferation (CCK-8/EdU), migration, and invasion (Transwell) post-WISP1 silencing (scale bars = 50μm). (M) ELISA quantification of secreted WISP1 in supernatants from untransfected CAFs, CAFs-shVector, and CAFs-shWISP1 at 0 h, 24 h, and 48 h (N) Schematic of indirect co-culture system for CAF-ESCC interaction analysis. (L, P) WB assessment of extracellular matrix (ECM)-remodeling markers (COL1A1, MMP14) in WISP1-depleted CAFs and rescue via recombinant human WISP1 (rhWISP1). (O, Q) Transwell assay comparing the migration and invasion capacities of KYSE150 (left) and Eca109 (right) cells co-cultured with untransfected CAFs, CAFs-shVector, or CAFs-shWISP1(scale bars = 50μm). (R, S) Transwell assay assessing the migration and invasion capacities of KYSE150 (left) and Eca109 (right) cells in the CAFs-shWISP1 co-culture system following rescue experiments with rhWISP1 supplementation (scale bar = 50μm).Data are presented as mean ± standard deviation from three independent experiments. Statistical significance was analyzed by two-tailed Student’s t-tests. ns = not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: WISP1 regulates cancer-associated fibroblast function and extracellular matrix remodeling. (A, B) Immunofluorescence multiplex staining of α-smooth muscle actin (αSMA) and fibroblast activation protein (FAP) in paired cancer-associated fibroblasts (CAFs, tumor-derived) and normal fibroblasts (NFs, adjacent non-tumor tissues) (scale bars = 50μm). (C) Western blot (WB) analysis of WISP1, αSMA, and FAP expression in CAFs versus NFs. (D, E) Lentiviral short hairpin RNA (shRNA)-mediated WISP1 knockdown in CAFs, validated by quantitative reverse transcription PCR (qRT-PCR) and WB. (F–K) Functional characterization of CAF proliferation (CCK-8/EdU), migration, and invasion (Transwell) post-WISP1 silencing (scale bars = 50μm). (M) ELISA quantification of secreted WISP1 in supernatants from untransfected CAFs, CAFs-shVector, and CAFs-shWISP1 at 0 h, 24 h, and 48 h (N) Schematic of indirect co-culture system for CAF-ESCC interaction analysis. (L, P) WB assessment of extracellular matrix (ECM)-remodeling markers (COL1A1, MMP14) in WISP1-depleted CAFs and rescue via recombinant human WISP1 (rhWISP1). (O, Q) Transwell assay comparing the migration and invasion capacities of KYSE150 (left) and Eca109 (right) cells co-cultured with untransfected CAFs, CAFs-shVector, or CAFs-shWISP1(scale bars = 50μm). (R, S) Transwell assay assessing the migration and invasion capacities of KYSE150 (left) and Eca109 (right) cells in the CAFs-shWISP1 co-culture system following rescue experiments with rhWISP1 supplementation (scale bar = 50μm).Data are presented as mean ± standard deviation from three independent experiments. Statistical significance was analyzed by two-tailed Student’s t-tests. ns = not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Techniques: Immunofluorescence, Multiplex Assay, Staining, Activation Assay, Derivative Assay, Western Blot, Expressing, shRNA, Knockdown, Reverse Transcription, Quantitative RT-PCR, Functional Assay, CCK-8 Assay, Migration, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Recombinant, Transwell Assay, Cell Culture, Standard Deviation, Two Tailed Test

WISP1 regulates ECM remodeling in CAFs through STAT3 signaling. (A, B) Representative phospho-kinase antibody array membrane comparing CAFs-shVector and CAFs-shWISP1. Red boxes highlight phosphorylated STAT3 (Y705) signals. (C) Dose-response curve of STAT3 inhibitor Stattic in CAFs. (D) Western blot analysis of phosphorylated STAT3 (Y705), total STAT3, and GAPDH (loading control) in: CAFs-shVector, CAFs-shWISP1, CAFs-shWISP1 + rhWISP1 (0.8 μg/mL), and CAFs-shWISP1 + rhWISP1 (0.8 μg/mL) + Stattic (7 μM). (E) Western blot analysis of COL1A1, MMP14, and GAPDH in CAFs-shWISP1 + rhWISP1 (0.8 μg/mL) and CAFs-shWISP1 + rhWISP1 (0.8 μg/mL) + Stattic (7 μM).

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: WISP1 regulates ECM remodeling in CAFs through STAT3 signaling. (A, B) Representative phospho-kinase antibody array membrane comparing CAFs-shVector and CAFs-shWISP1. Red boxes highlight phosphorylated STAT3 (Y705) signals. (C) Dose-response curve of STAT3 inhibitor Stattic in CAFs. (D) Western blot analysis of phosphorylated STAT3 (Y705), total STAT3, and GAPDH (loading control) in: CAFs-shVector, CAFs-shWISP1, CAFs-shWISP1 + rhWISP1 (0.8 μg/mL), and CAFs-shWISP1 + rhWISP1 (0.8 μg/mL) + Stattic (7 μM). (E) Western blot analysis of COL1A1, MMP14, and GAPDH in CAFs-shWISP1 + rhWISP1 (0.8 μg/mL) and CAFs-shWISP1 + rhWISP1 (0.8 μg/mL) + Stattic (7 μM).

Techniques: Ab Array, Membrane, Western Blot, Control

Journal: bioRxiv

Article Title: WISP1 drives a mechanically active immune modulatory and proliferative cardiac myofibroblast state

doi: 10.64898/2026.02.17.706476

Figure Lengend Snippet: (A) Wisp1 mRNA expression in primary adult mouse cardiac fibroblasts following TGFβ1 (10 ng/mL) treatment for 24–72 h. (B) Representative immunofluorescence images of α-SMA (green), vimentin (red), and DAPI (blue) after 72 h treatment with vehicle (Ctrl), WISP1 (500 ng/mL), TGFβ1 (10 ng/mL), or WISP1 + TGFβ1. (C, D) Representative Western blot (C) and densitometric quantification (D) of α-SMA normalized to GAPDH. Data are mean ± SEM from ≥3 independent isolations. Statistical analysis by one-way ANOVA with Tukey’s post hoc test; *p < 0.05. Data are mean ± SEM from ≥3 independent isolations. Statistical analysis by one-way ANOVA with Tukey’s post hoc test; *p < 0.05.

Article Snippet: For treatments, CFs were seeded in 12- or 24-well plates in DMEM + 10% FBS and treated for 72 h with recombinant human TGFβ1 (10 ng/mL; R&D Systems, 7754-BH) and/or recombinant murine WISP1 (500 ng/mL; R&D Systems, 1680-WS-050).

Techniques: Expressing, Immunofluorescence, Western Blot

Journal: bioRxiv

Article Title: WISP1 drives a mechanically active immune modulatory and proliferative cardiac myofibroblast state

doi: 10.64898/2026.02.17.706476

Figure Lengend Snippet: Primary adult mouse cardiac fibroblasts were embedded in floating collagen gels and treated for 24 h with vehicle (Ctrl), WISP1 (500 ng/mL), TGFβ1 (10 ng/mL), or WISP1 + TGFβ1. (A) Representative gel images at 0 h and 24 h. (B) Quantification of gel area (% contraction; n = 4 isolations, 2 males & 2 females). (C–D) Confluent fibroblast monolayers were scratched and imaged every 2 hours for 24 hours using the same treatment conditions. (C) Representative bright-field images at time point 0 and 14. (D) Quantification of wound closure (% of gap closed relative to time 0; n = 8 isolations, 4 male & 4 female). Data are mean ± SEM. One-way ANOVA with Tukey’s multiple comparisons; *p ≤ 0.05.

Techniques:

Figure 3. Knockdown of WPMY-1 suppresses LIF-driven neuroendocrine differentiation (NED) and malignant progression in prostate cancer (PCa) cells (A) Relative protein levels of LIF, LIFR, WISP1, SOCS3, and PDL1 were measured in AR-positive PCa cell lines (LNCaP, C4-2, and 22Rv1), an AR-negative PCa cell line (PC3), and an NEPC cell line (LASCPC-01). (B) Relative protein levels of WISP1, phosphorylated (p)-STAT3, STAT3, PDL1, and SOCS3 were measured in LNCaP cells treated with PBS or 100 ng/mL of the LIF recombinant protein for a duration ranging 0 to 96 h. (C) Relative mRNA levels of WISP1, neuroendocrine (CHGA, SYP, and ENO2), stem cell (SOX2 and NANOG), and anti-inﬂammatory (SOCS3 and PDL1) markers in LNCaP cells treated with PBS or 100 ng/mL of the LIF recombinant protein or combined treatment with DMSO or 35 nM EC330 for 48 h * vs. PBS+DMSO; # vs. LIF+DMSO, as determined by a one-way ANOVA. (D) Relative protein levels of WISP1, p-STAT3, STAT3, PDL1, and SOCS3 in LNCaP and C4-2 cells cultured in PBS or 100 ng/mL of the LIF recombinant protein, followed by DMSO or 35 nM EC330 treatment for 48 h. (E) Relative mRNA expression levels of WISP1, neuroendocrine markers, stem cell markers, and anti-inﬂammatory markers (SOCS3 and PDL1) were measured in LNCaP cells expressing the non-target control (NC) or WISP1 siRNA, followed by treatment with PBS or 100 ng/mL of the LIF recombinant protein for 48 h * vs. PBS+NC; # vs. LIF+NC, as determined by a one-way ANOVA. (F and G) Relative cell proliferation (F) and sphere formation (G) were measured in LNCaP cells expressing either NC or WISP1 siRNA, followed by treatment with either PBS or 100 ng/mL of LIF recombinant protein for 5 days (F) or 1 week (G). Scale bars represent 100 mm (G). * vs. PBS+NC; # vs. LIF+NC, as determined by a one-way ANOVA. (H and I) Relative cell migration (H) and invasion through Matrigel (I) were measured in LNCaP cells expressing the NC or WISP1 siRNA, followed by treatment with PBS or 100 ng/mL of LIF recombinant protein for 12 h. Scale bars representing 20 mm are shown. * vs. PBS+NC; # vs. LIF+NC, as determined by a one-way ANOVA. Quantiﬁcation of relative mRNA levels, cell proliferation, sphere formation, and cell invasion through Matrigel is presented as the mean G SEM from three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.005. (J) A GSEA of the TCGA PCa dataset revealed signiﬁcant associations between high WISP1 expression in prostate tissues and a gene signature representing NEPC-responsive signaling. NES, normalized enrichment score; FDR, false discovery rate.

Journal: iScience

Article Title: WNT1-inducible signaling pathway protein 1 activation through C-X-C motif chemokine ligand 5/C-X-C chemokine receptor type 2/leukemia inhibitory factor/leukemia inhibitory factor receptor signaling promotes immunosuppression and neuroendocrine differentiation in prostate cancer.

doi: 10.1016/j.isci.2024.110562

Figure Lengend Snippet: Figure 3. Knockdown of WPMY-1 suppresses LIF-driven neuroendocrine differentiation (NED) and malignant progression in prostate cancer (PCa) cells (A) Relative protein levels of LIF, LIFR, WISP1, SOCS3, and PDL1 were measured in AR-positive PCa cell lines (LNCaP, C4-2, and 22Rv1), an AR-negative PCa cell line (PC3), and an NEPC cell line (LASCPC-01). (B) Relative protein levels of WISP1, phosphorylated (p)-STAT3, STAT3, PDL1, and SOCS3 were measured in LNCaP cells treated with PBS or 100 ng/mL of the LIF recombinant protein for a duration ranging 0 to 96 h. (C) Relative mRNA levels of WISP1, neuroendocrine (CHGA, SYP, and ENO2), stem cell (SOX2 and NANOG), and anti-inﬂammatory (SOCS3 and PDL1) markers in LNCaP cells treated with PBS or 100 ng/mL of the LIF recombinant protein or combined treatment with DMSO or 35 nM EC330 for 48 h * vs. PBS+DMSO; # vs. LIF+DMSO, as determined by a one-way ANOVA. (D) Relative protein levels of WISP1, p-STAT3, STAT3, PDL1, and SOCS3 in LNCaP and C4-2 cells cultured in PBS or 100 ng/mL of the LIF recombinant protein, followed by DMSO or 35 nM EC330 treatment for 48 h. (E) Relative mRNA expression levels of WISP1, neuroendocrine markers, stem cell markers, and anti-inﬂammatory markers (SOCS3 and PDL1) were measured in LNCaP cells expressing the non-target control (NC) or WISP1 siRNA, followed by treatment with PBS or 100 ng/mL of the LIF recombinant protein for 48 h * vs. PBS+NC; # vs. LIF+NC, as determined by a one-way ANOVA. (F and G) Relative cell proliferation (F) and sphere formation (G) were measured in LNCaP cells expressing either NC or WISP1 siRNA, followed by treatment with either PBS or 100 ng/mL of LIF recombinant protein for 5 days (F) or 1 week (G). Scale bars represent 100 mm (G). * vs. PBS+NC; # vs. LIF+NC, as determined by a one-way ANOVA. (H and I) Relative cell migration (H) and invasion through Matrigel (I) were measured in LNCaP cells expressing the NC or WISP1 siRNA, followed by treatment with PBS or 100 ng/mL of LIF recombinant protein for 12 h. Scale bars representing 20 mm are shown. * vs. PBS+NC; # vs. LIF+NC, as determined by a one-way ANOVA. Quantiﬁcation of relative mRNA levels, cell proliferation, sphere formation, and cell invasion through Matrigel is presented as the mean G SEM from three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.005. (J) A GSEA of the TCGA PCa dataset revealed signiﬁcant associations between high WISP1 expression in prostate tissues and a gene signature representing NEPC-responsive signaling. NES, normalized enrichment score; FDR, false discovery rate.

Article Snippet: Protein levels of WISP1 were quantified using a human WISP1 ELISA kit (Elabscience, #E-EL-H5542) following the manufacturer’s instructions.

Techniques: Knockdown, Recombinant, Cell Culture, Expressing, Control, Migration

Figure 4. Crosstalk between prostate cancer (PCa) cells and prostate stromal cells promotes neuroendocrine differentiation (NED) and expressions of immunosuppressive cytokines in the tumor microenvironment (TME) through the activation of LIF/LIFR signaling (A) Relative mRNA expression levels of WISP1, neuroendocrine markers (CHGA, SYP, and ENO2), stem cell markers (SOX2 and NANOG), anti-inﬂammatory markers (SOCS3 and PDL1), LIF, CXCR2, and CXCL5 in LNCaP cells. These cells were cultured with conditioned medium (CM) collected from human WPMY-1 stromal cells, at concentrations of 0%, 15%, or 50%, for a duration of 48 h * vs. 0%, as determined by a one-way ANOVA. (B) Relative mRNA expression levels of WISP1, neuroendocrine markers, stem cell markers, anti-inﬂammatory markers, LIF, CXCR2, and CXCL5 were measured in LNCaP cells. These cells were cultured with CM obtained from WPMY-1 stromal cells treated with DMSO or 35 nM EC330 for 48 h * vs. Veh+DMSO; # vs. WPMY-1 CM + DMSO, as determined by a one-way ANOVA. (C) Relative mRNA expression levels of WISP1, neuroendocrine, stem cell, anti-inﬂammatory markers, LIF, CXCR2, and CXCL5 in LNCaP cells cultured with CM collected from the non-target control (Luc) or LIFR shRNA-expressing WPMY-1 cells for 48 h * vs. Veh; # vs. WPMY-1/shLuc CM, as determined by a one-way ANOVA. (D) Relative mRNA expression levels of LIFR, LIF, CXCL5, WISP1, and anti-inﬂammatory markers (IL10, IL4, IL1RN, TGFB1, VEGFA, IFNA17, and SOCS3) in WPMY-1 cells stably expressing the shLuc or LIFR shRNA. * vs. shLuc, as determined by a one-way ANOVA. (E) Relative mRNA expression levels of LIFR, LIF, CXCL5, WISP1, and anti-inﬂammatory markers in WPMY-1 cells treated with PBS or 100 ng/mL of the LIF recombinant protein, followed by DMSO or 35 nM EC330 treatment for 48 h * vs. PBS+DMSO; # vs. LIF+DMSO, as determined by a one-way ANOVA. (F and G) Cell proliferation (F) and sphere formation (G) of LNCaP cells were evaluated. These cells were cultured with CM obtained from WPMY-1 stromal cells treated with DMOS or 35 nM EC330 for 48 h. Scale bars in (G) represent 100 mm. Statistical comparisons were performed using a one-way ANOVA. * vs. Veh+DMSO; # vs. WPMY-1 CM + DMSO. (H and I) Relative cell migration (H) and invasion through Matrigel (I) were measured in LNCaP cells. These cells were cultured with CM obtained from WPMY-1 stromal cells treated with DMSO or 35 nM EC330 for 12 h. Scale bars representing 20 mm are shown. * vs. Veh+DMSO; # vs. WPMY-1 CM + DMSO, as determined by a one-way ANOVA. Quantiﬁcation of relative mRNA levels, cell proliferation, sphere formation, and cell invasion through Matrigel is presented as the mean G SEM, based on three biological replicates. Signiﬁcance levels are denoted as *p < 0.05, **p < 0.01, and ***p < 0.005.

Journal: iScience

doi: 10.1016/j.isci.2024.110562

Figure Lengend Snippet: Figure 4. Crosstalk between prostate cancer (PCa) cells and prostate stromal cells promotes neuroendocrine differentiation (NED) and expressions of immunosuppressive cytokines in the tumor microenvironment (TME) through the activation of LIF/LIFR signaling (A) Relative mRNA expression levels of WISP1, neuroendocrine markers (CHGA, SYP, and ENO2), stem cell markers (SOX2 and NANOG), anti-inﬂammatory markers (SOCS3 and PDL1), LIF, CXCR2, and CXCL5 in LNCaP cells. These cells were cultured with conditioned medium (CM) collected from human WPMY-1 stromal cells, at concentrations of 0%, 15%, or 50%, for a duration of 48 h * vs. 0%, as determined by a one-way ANOVA. (B) Relative mRNA expression levels of WISP1, neuroendocrine markers, stem cell markers, anti-inﬂammatory markers, LIF, CXCR2, and CXCL5 were measured in LNCaP cells. These cells were cultured with CM obtained from WPMY-1 stromal cells treated with DMSO or 35 nM EC330 for 48 h * vs. Veh+DMSO; # vs. WPMY-1 CM + DMSO, as determined by a one-way ANOVA. (C) Relative mRNA expression levels of WISP1, neuroendocrine, stem cell, anti-inﬂammatory markers, LIF, CXCR2, and CXCL5 in LNCaP cells cultured with CM collected from the non-target control (Luc) or LIFR shRNA-expressing WPMY-1 cells for 48 h * vs. Veh; # vs. WPMY-1/shLuc CM, as determined by a one-way ANOVA. (D) Relative mRNA expression levels of LIFR, LIF, CXCL5, WISP1, and anti-inﬂammatory markers (IL10, IL4, IL1RN, TGFB1, VEGFA, IFNA17, and SOCS3) in WPMY-1 cells stably expressing the shLuc or LIFR shRNA. * vs. shLuc, as determined by a one-way ANOVA. (E) Relative mRNA expression levels of LIFR, LIF, CXCL5, WISP1, and anti-inﬂammatory markers in WPMY-1 cells treated with PBS or 100 ng/mL of the LIF recombinant protein, followed by DMSO or 35 nM EC330 treatment for 48 h * vs. PBS+DMSO; # vs. LIF+DMSO, as determined by a one-way ANOVA. (F and G) Cell proliferation (F) and sphere formation (G) of LNCaP cells were evaluated. These cells were cultured with CM obtained from WPMY-1 stromal cells treated with DMOS or 35 nM EC330 for 48 h. Scale bars in (G) represent 100 mm. Statistical comparisons were performed using a one-way ANOVA. * vs. Veh+DMSO; # vs. WPMY-1 CM + DMSO. (H and I) Relative cell migration (H) and invasion through Matrigel (I) were measured in LNCaP cells. These cells were cultured with CM obtained from WPMY-1 stromal cells treated with DMSO or 35 nM EC330 for 12 h. Scale bars representing 20 mm are shown. * vs. Veh+DMSO; # vs. WPMY-1 CM + DMSO, as determined by a one-way ANOVA. Quantiﬁcation of relative mRNA levels, cell proliferation, sphere formation, and cell invasion through Matrigel is presented as the mean G SEM, based on three biological replicates. Signiﬁcance levels are denoted as *p < 0.05, **p < 0.01, and ***p < 0.005.

Article Snippet: Protein levels of WISP1 were quantified using a human WISP1 ELISA kit (Elabscience, #E-EL-H5542) following the manufacturer’s instructions.

Techniques: Activation Assay, Expressing, Cell Culture, Control, shRNA, Stable Transfection, Recombinant, Migration

Figure 5. LIF/STAT3-driven transcription of WISP1 through direct binding to the gamma interferon activation site (GAS) of the regulatory sequence (A) ChIP-sequencing analysis was performed to detect the GAS for WISP1. Detected GAS sites are labeled as black boxes in the tracks. ChIP-sequencing data were downloaded from Gene Expression Omnibus (GEO) (GSM2752900) and analyzed using the Genome Browser (Genomics Institute, UCSC). (B) A schematic representation is shown for the predicted wild-type (WT) and mutant (M)-GASs in the regulatory sequence reporter constructs of the human WISP1 gene (GRCh38:8) (C and D) A ChIP assay was performed to show the binding of phosphorylated (p)-STAT3 to the predicted GAS in the WISP1 gene regulatory sequence. The assay was conducted in LNCaP cells (C) or WPMY-1 cells (D) treated with PBS or 100 ng/mL of the LIF recombinant protein, followed by treatment with DMSO or with 10 or 35 nM EC330 for 48 h. Sheared chromatin from nuclear extracts was precipitated with antibodies to p-STAT3 or control IgG, and predictive primers (B, indicated by black arrows) were used to quantify the precipitated DNA using a qPCR. Enrichment of each protein at each site is presented as a percentage of the total input and then normalized to IgG. * vs. PBS+DMSO; # vs. LIF+DMSO, as determined by a one-way ANOVA. (E) A ChIP assay was performed to demonstrate the binding of p-STAT3 to the predicted GAS in the regulatory sequence of the LIF gene in WPMY-1 cells. Cells were treated with PBS or 100 ng/mL of the LIF recombinant protein, followed by treatment with DMSO or with 10 or 35 nM EC330 for 48 h * vs. PBS+DMSO; # vs. LIF+DMSO, as determined by a one-way ANOVA. (F–I) The relative mean ﬂuorescence intensity (MFI) of the GFP reporter gene containing either the WT-GAS or M-GAS from the WISP1 regulatory sequence was measured in LNCaP cells (F and G) and WPMY-1 cells (H and I) after treatment with PBS or 100 ng/mL of LIF recombinant protein. This was followed by further treatment with either DMSO or EC330 (10 or 35 nM) for 48 h (F and H). * vs. WT/PBS+DMSO (F and H) or WT + PBS (G and I); # vs. WT/LIF+DMSO (F and H) or WT + LIF (G and I), as determined by a one-way ANOVA. (J and K) The relative MFI of the GFP reporter gene containing the WT-GAS or M-GAS from the LIF regulatory sequence was measured in WPMY-1 cells treated with PBS or 100 ng/mL of LIF recombinant protein or combined treatment with DMSO or 10 or 35 nM EC330 for 48 h * vs. WT/PBS+DMSO (J) or WT + PBS (K); # vs. WT/LIF+DMSO (J) or WT + LIF (K), as determined by a one-way ANOVA. Quantiﬁcation of relative p-STAT3 enrichment and MFI is presented as the mean G SEM from three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.005.

Journal: iScience

doi: 10.1016/j.isci.2024.110562

Figure Lengend Snippet: Figure 5. LIF/STAT3-driven transcription of WISP1 through direct binding to the gamma interferon activation site (GAS) of the regulatory sequence (A) ChIP-sequencing analysis was performed to detect the GAS for WISP1. Detected GAS sites are labeled as black boxes in the tracks. ChIP-sequencing data were downloaded from Gene Expression Omnibus (GEO) (GSM2752900) and analyzed using the Genome Browser (Genomics Institute, UCSC). (B) A schematic representation is shown for the predicted wild-type (WT) and mutant (M)-GASs in the regulatory sequence reporter constructs of the human WISP1 gene (GRCh38:8) (C and D) A ChIP assay was performed to show the binding of phosphorylated (p)-STAT3 to the predicted GAS in the WISP1 gene regulatory sequence. The assay was conducted in LNCaP cells (C) or WPMY-1 cells (D) treated with PBS or 100 ng/mL of the LIF recombinant protein, followed by treatment with DMSO or with 10 or 35 nM EC330 for 48 h. Sheared chromatin from nuclear extracts was precipitated with antibodies to p-STAT3 or control IgG, and predictive primers (B, indicated by black arrows) were used to quantify the precipitated DNA using a qPCR. Enrichment of each protein at each site is presented as a percentage of the total input and then normalized to IgG. * vs. PBS+DMSO; # vs. LIF+DMSO, as determined by a one-way ANOVA. (E) A ChIP assay was performed to demonstrate the binding of p-STAT3 to the predicted GAS in the regulatory sequence of the LIF gene in WPMY-1 cells. Cells were treated with PBS or 100 ng/mL of the LIF recombinant protein, followed by treatment with DMSO or with 10 or 35 nM EC330 for 48 h * vs. PBS+DMSO; # vs. LIF+DMSO, as determined by a one-way ANOVA. (F–I) The relative mean ﬂuorescence intensity (MFI) of the GFP reporter gene containing either the WT-GAS or M-GAS from the WISP1 regulatory sequence was measured in LNCaP cells (F and G) and WPMY-1 cells (H and I) after treatment with PBS or 100 ng/mL of LIF recombinant protein. This was followed by further treatment with either DMSO or EC330 (10 or 35 nM) for 48 h (F and H). * vs. WT/PBS+DMSO (F and H) or WT + PBS (G and I); # vs. WT/LIF+DMSO (F and H) or WT + LIF (G and I), as determined by a one-way ANOVA. (J and K) The relative MFI of the GFP reporter gene containing the WT-GAS or M-GAS from the LIF regulatory sequence was measured in WPMY-1 cells treated with PBS or 100 ng/mL of LIF recombinant protein or combined treatment with DMSO or 10 or 35 nM EC330 for 48 h * vs. WT/PBS+DMSO (J) or WT + PBS (K); # vs. WT/LIF+DMSO (J) or WT + LIF (K), as determined by a one-way ANOVA. Quantiﬁcation of relative p-STAT3 enrichment and MFI is presented as the mean G SEM from three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.005.

Article Snippet: Protein levels of WISP1 were quantified using a human WISP1 ELISA kit (Elabscience, #E-EL-H5542) following the manufacturer’s instructions.

Techniques: Binding Assay, Activation Assay, Sequencing, ChIP-sequencing, Labeling, Gene Expression, Mutagenesis, Construct, Recombinant, Control

Figure 6. WISP1 abundance in serum relative to prostate cancer (PCa) progression (A) Relative mRNA levels of CXCL5, CXCR2, LIF, WISP1, neuroendocrine (CHGA, SYP, and ENO2), and stem cell (SOX2 and NANOG) markers in C4-2 cells expressing the empty vector (EV) or CXCL5-expressing vector, followed by treatment with DMSO or 35 nM EC330 for 48 h * vs. EV + DMSO; # vs. CXCL5+DMSO, as determined by a one-way ANOVA. (B and C) Tumor growth analysis was conducted by subcutaneously inoculating male nude mice with C4-2 cells expressing either the EV or a CXCL5-expressing vector. The mice were then treated bi-daily with either DMSO or 2.5 mg/kg EC330 via intraperitoneal injection and allowed to grow for 8 weeks. Tumor sizes were measured weekly (B). Tumor weights were measured upon tumor collection (C). n = 5 per group. * vs. EV + DMSO; # vs. CXCL5+DMSO, as determined by a one- way ANOVA and t-test. (D and E) Immunohistochemical (IHC) staining and intensity analyses were performed to assess protein levels of CXCL5, CXCR2, LIF, WISP1, and ENO2 in subcutaneous tumors derived from (B). * vs. EV + DMSO; # vs. CXCL5+DMSO, as determined by a two-tailed Student’s t-test. Scale bars, 100 mm. (F) WISP1 concentrations were measured in patient sera derived from samples of benign prostatic hyperplasia (BPH; n = 10), hormone-sensitive PCa (HSPC, n = 10), and metastatic castration-resistant PCa (mCRPC; n = 8). * vs. BPH; # vs. HSPC, analyzed by a one-way ANOVA.

Journal: iScience

doi: 10.1016/j.isci.2024.110562

Figure Lengend Snippet: Figure 6. WISP1 abundance in serum relative to prostate cancer (PCa) progression (A) Relative mRNA levels of CXCL5, CXCR2, LIF, WISP1, neuroendocrine (CHGA, SYP, and ENO2), and stem cell (SOX2 and NANOG) markers in C4-2 cells expressing the empty vector (EV) or CXCL5-expressing vector, followed by treatment with DMSO or 35 nM EC330 for 48 h * vs. EV + DMSO; # vs. CXCL5+DMSO, as determined by a one-way ANOVA. (B and C) Tumor growth analysis was conducted by subcutaneously inoculating male nude mice with C4-2 cells expressing either the EV or a CXCL5-expressing vector. The mice were then treated bi-daily with either DMSO or 2.5 mg/kg EC330 via intraperitoneal injection and allowed to grow for 8 weeks. Tumor sizes were measured weekly (B). Tumor weights were measured upon tumor collection (C). n = 5 per group. * vs. EV + DMSO; # vs. CXCL5+DMSO, as determined by a one- way ANOVA and t-test. (D and E) Immunohistochemical (IHC) staining and intensity analyses were performed to assess protein levels of CXCL5, CXCR2, LIF, WISP1, and ENO2 in subcutaneous tumors derived from (B). * vs. EV + DMSO; # vs. CXCL5+DMSO, as determined by a two-tailed Student’s t-test. Scale bars, 100 mm. (F) WISP1 concentrations were measured in patient sera derived from samples of benign prostatic hyperplasia (BPH; n = 10), hormone-sensitive PCa (HSPC, n = 10), and metastatic castration-resistant PCa (mCRPC; n = 8). * vs. BPH; # vs. HSPC, analyzed by a one-way ANOVA.

Article Snippet: Protein levels of WISP1 were quantified using a human WISP1 ELISA kit (Elabscience, #E-EL-H5542) following the manufacturer’s instructions.

Techniques: Expressing, Plasmid Preparation, Injection, Immunohistochemical staining, Immunohistochemistry, Derivative Assay, Two Tailed Test

( A ) F4/80 positive cells were accumulated in mice kidney (kidney cortex) with WISP1 administration and reduced by PTL, which was verified by counting the percentage of F4/80 positive cells. ( B ) The expression of inflammation markers was up-regulated by WISP1 and alleviated by PTL treatment. ( C ) M1 and M2 marker expression. Results are expressed as the mean ± SEM relative gene expression, *** P <0.001,** P <0.01, * P <0.05. n =3–5.

Journal: Clinical Science (London, England : 1979)

Article Title: WNT1-inducible signaling pathway protein 1 regulates kidney inflammation through the NF-κB pathway

doi: 10.1042/CS20210663

Figure Lengend Snippet: ( A ) F4/80 positive cells were accumulated in mice kidney (kidney cortex) with WISP1 administration and reduced by PTL, which was verified by counting the percentage of F4/80 positive cells. ( B ) The expression of inflammation markers was up-regulated by WISP1 and alleviated by PTL treatment. ( C ) M1 and M2 marker expression. Results are expressed as the mean ± SEM relative gene expression, *** P <0.001,** P <0.01, * P <0.05. n =3–5.

Article Snippet: Recombinant human WISP1 protein, IgG control and mouse WISP1 antibody were purchased commercially (R&D Systems, Minneapolis, MN) and used at specified concentrations.

Techniques: Expressing, Marker, Gene Expression

At 48 h after administration of WISP1 and/or PTL to healthy mice, there were: ( A ) no morphology changes (Masson staining) observed based on in the kidney at 48 h after administration of WISP1 and/or PTL in healthy mice. ( B ) No collagen (PSR staining) alteration was found in the WISP1-treated mice. ( C ) Q-PCR data showed no significant changes in collagen1a1 and fibronectin. ( D ) α-SMA expression were up-regulated with WISP1 treatment, reduced by PTL. Results are expressed as the mean ± SEM relative gene expression, ** P <0.01, * P <0.05. n =3–5.

Journal: Clinical Science (London, England : 1979)

Article Title: WNT1-inducible signaling pathway protein 1 regulates kidney inflammation through the NF-κB pathway

doi: 10.1042/CS20210663

Figure Lengend Snippet: At 48 h after administration of WISP1 and/or PTL to healthy mice, there were: ( A ) no morphology changes (Masson staining) observed based on in the kidney at 48 h after administration of WISP1 and/or PTL in healthy mice. ( B ) No collagen (PSR staining) alteration was found in the WISP1-treated mice. ( C ) Q-PCR data showed no significant changes in collagen1a1 and fibronectin. ( D ) α-SMA expression were up-regulated with WISP1 treatment, reduced by PTL. Results are expressed as the mean ± SEM relative gene expression, ** P <0.01, * P <0.05. n =3–5.

Article Snippet: Recombinant human WISP1 protein, IgG control and mouse WISP1 antibody were purchased commercially (R&D Systems, Minneapolis, MN) and used at specified concentrations.

Techniques: Staining, Expressing, Gene Expression

( A ) The translocation of P-p65 into nucleus induced by WISP1 was inhibited by treatment of NFκB inhibitor PTL, with the inhibition on the up-regulated expression of TNF-α. ( B , C ) PTL inhibited the LPS-induced nucleus translocation of P-p65 and the up-regulated expression of inflammation markers. ( D ) Results are expressed as the mean ± SEM relative gene expression, *** P <0.001,** P <0.01. n =3–5.

Journal: Clinical Science (London, England : 1979)

Article Title: WNT1-inducible signaling pathway protein 1 regulates kidney inflammation through the NF-κB pathway

doi: 10.1042/CS20210663

Figure Lengend Snippet: ( A ) The translocation of P-p65 into nucleus induced by WISP1 was inhibited by treatment of NFκB inhibitor PTL, with the inhibition on the up-regulated expression of TNF-α. ( B , C ) PTL inhibited the LPS-induced nucleus translocation of P-p65 and the up-regulated expression of inflammation markers. ( D ) Results are expressed as the mean ± SEM relative gene expression, *** P <0.001,** P <0.01. n =3–5.

Article Snippet: Recombinant human WISP1 protein, IgG control and mouse WISP1 antibody were purchased commercially (R&D Systems, Minneapolis, MN) and used at specified concentrations.

Techniques: Translocation Assay, Inhibition, Expressing, Gene Expression

( A ) Knockdown (KD) of WISP1 by shRNA. ( B ) Knockdown of WISP1 in LPS-stimulated RAW macrophages inhibited ( B ) the translocation of P-p65 into nucleus, and up-regulated ( C ) expression of inflammation markers. *** P <0.001,** P <0.01. n =3–5.

Journal: Clinical Science (London, England : 1979)

Article Title: WNT1-inducible signaling pathway protein 1 regulates kidney inflammation through the NF-κB pathway

doi: 10.1042/CS20210663

Figure Lengend Snippet: ( A ) Knockdown (KD) of WISP1 by shRNA. ( B ) Knockdown of WISP1 in LPS-stimulated RAW macrophages inhibited ( B ) the translocation of P-p65 into nucleus, and up-regulated ( C ) expression of inflammation markers. *** P <0.001,** P <0.01. n =3–5.

Article Snippet: Recombinant human WISP1 protein, IgG control and mouse WISP1 antibody were purchased commercially (R&D Systems, Minneapolis, MN) and used at specified concentrations.

Techniques: Knockdown, shRNA, Translocation Assay, Expressing

( A ) The addition of WISP1 antibody (Ab) inhibited the translocation of P-p65 induced by LPS and the expression of inflammation markers ( B,C ). ELISA showed that the up-regulation TNF-α was reduced by WISP1 knockdown and the addition of antibody. Results are expressed as the mean ± SEM relative gene expression, *** P <0.001,** P <0.01, * P <0.05. n =3–5.

Journal: Clinical Science (London, England : 1979)

Article Title: WNT1-inducible signaling pathway protein 1 regulates kidney inflammation through the NF-κB pathway

doi: 10.1042/CS20210663

Figure Lengend Snippet: ( A ) The addition of WISP1 antibody (Ab) inhibited the translocation of P-p65 induced by LPS and the expression of inflammation markers ( B,C ). ELISA showed that the up-regulation TNF-α was reduced by WISP1 knockdown and the addition of antibody. Results are expressed as the mean ± SEM relative gene expression, *** P <0.001,** P <0.01, * P <0.05. n =3–5.

Article Snippet: Recombinant human WISP1 protein, IgG control and mouse WISP1 antibody were purchased commercially (R&D Systems, Minneapolis, MN) and used at specified concentrations.

Techniques: Translocation Assay, Expressing, Enzyme-linked Immunosorbent Assay, Knockdown, Gene Expression

The representative cell images were taken using 4× light microscope. ( A ) PTL inhibited kidney fibroblast cells proliferation induced by recombinant WISP1 protein (1 μg/ml), validated by counting cell numbers. ( B ) Wisp1 knocked-down using shRNA reduced the proliferation of kidney fibroblast cells by TNF-α, confirmed by counting cell number. ( C ) The addition of WISP1 antibody reduced the fibroblast cell growth in the presence of TNF-α, as confirmed by counting cell number. n =3.

Journal: Clinical Science (London, England : 1979)

Article Title: WNT1-inducible signaling pathway protein 1 regulates kidney inflammation through the NF-κB pathway

doi: 10.1042/CS20210663

Figure Lengend Snippet: The representative cell images were taken using 4× light microscope. ( A ) PTL inhibited kidney fibroblast cells proliferation induced by recombinant WISP1 protein (1 μg/ml), validated by counting cell numbers. ( B ) Wisp1 knocked-down using shRNA reduced the proliferation of kidney fibroblast cells by TNF-α, confirmed by counting cell number. ( C ) The addition of WISP1 antibody reduced the fibroblast cell growth in the presence of TNF-α, as confirmed by counting cell number. n =3.

Article Snippet: Recombinant human WISP1 protein, IgG control and mouse WISP1 antibody were purchased commercially (R&D Systems, Minneapolis, MN) and used at specified concentrations.

Techniques: Light Microscopy, Recombinant, shRNA

( A ) STZ-induced DN mice at 18 weeks demonstrated the increased expression of TNF-α, MCP-1, CD68, α-SMA and WISP1 gene by qPCR, compared with control mice ( n =12–15 mice/group). Immunofluorescence staining indicated the elevated expression of WISP1 in DN model. Macrophage co-localized with WISP1 staining (white arrow). ( B ) High glucose-treated RAW cells expressed more WISP1, TNF-α and MCP-1 than low glucose-treated cells. ( C ) The kidneys of mice with 7 days post-UUO surgery showed an increased expression of inflammation genes, TNF-α, MCP-1 and IL6 and an up-regulation of WISP1, compared with control kidneys ( n =5 mice/group). Immunofluorescence staining indicated the elevated expression of WISP1 in UUO model. Macrophage co-localized with WISP1 staining (white arrow). *** P <0.001,** P <0.01.

Journal: Clinical Science (London, England : 1979)

Article Title: WNT1-inducible signaling pathway protein 1 regulates kidney inflammation through the NF-κB pathway

doi: 10.1042/CS20210663

Figure Lengend Snippet: ( A ) STZ-induced DN mice at 18 weeks demonstrated the increased expression of TNF-α, MCP-1, CD68, α-SMA and WISP1 gene by qPCR, compared with control mice ( n =12–15 mice/group). Immunofluorescence staining indicated the elevated expression of WISP1 in DN model. Macrophage co-localized with WISP1 staining (white arrow). ( B ) High glucose-treated RAW cells expressed more WISP1, TNF-α and MCP-1 than low glucose-treated cells. ( C ) The kidneys of mice with 7 days post-UUO surgery showed an increased expression of inflammation genes, TNF-α, MCP-1 and IL6 and an up-regulation of WISP1, compared with control kidneys ( n =5 mice/group). Immunofluorescence staining indicated the elevated expression of WISP1 in UUO model. Macrophage co-localized with WISP1 staining (white arrow). *** P <0.001,** P <0.01.

Article Snippet: Recombinant human WISP1 protein, IgG control and mouse WISP1 antibody were purchased commercially (R&D Systems, Minneapolis, MN) and used at specified concentrations.

Techniques: Expressing, Control, Immunofluorescence, Staining

( A ) Q-PCR analysis demonstrated that WISP1 antibody reduced inflammation gene expression in kidneys from UUO mice. ( B ) UUO kidneys without treatment or receiving IgG show an up-regulated F4/80 positive cells accumulation at 7 days. Administration of WISP1 antibody in UUO mice resulted in a reduction, verified by the percentage of F4/80 positive cells. ( C ) WISP1 antibody treatment rescued the degradation of IκBα expression in UUO kidneys, by Western blotting. Results are expressed as the mean ± SEM relative gene expression, *** P <0.001, ** P <0.01, * P <0.05. n =3–7.

Journal: Clinical Science (London, England : 1979)

Article Title: WNT1-inducible signaling pathway protein 1 regulates kidney inflammation through the NF-κB pathway

doi: 10.1042/CS20210663

Figure Lengend Snippet: ( A ) Q-PCR analysis demonstrated that WISP1 antibody reduced inflammation gene expression in kidneys from UUO mice. ( B ) UUO kidneys without treatment or receiving IgG show an up-regulated F4/80 positive cells accumulation at 7 days. Administration of WISP1 antibody in UUO mice resulted in a reduction, verified by the percentage of F4/80 positive cells. ( C ) WISP1 antibody treatment rescued the degradation of IκBα expression in UUO kidneys, by Western blotting. Results are expressed as the mean ± SEM relative gene expression, *** P <0.001, ** P <0.01, * P <0.05. n =3–7.

Article Snippet: Recombinant human WISP1 protein, IgG control and mouse WISP1 antibody were purchased commercially (R&D Systems, Minneapolis, MN) and used at specified concentrations.

Techniques: Gene Expression, Expressing, Western Blot

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