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human wisp1 elisa kit  (BioVendor Instruments)


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    BioVendor Instruments human wisp1 elisa kit
    Human Wisp1 Elisa Kit, supplied by BioVendor Instruments, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+wisp1+elisa+kit/pmc09150549-107-6-10?v=BioVendor+Instruments
    Average 91 stars, based on 1 article reviews
    human wisp1 elisa kit - by Bioz Stars, 2026-07
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    <t>WISP1</t> is endogenously upregulated in primary human DHLF. WISP1 transcript and secreted protein are expressed in NHLF and DHLF. DHLF exhibit significantly higher ( A ) WISP1 mRNA transcript and ( B ) secreted WISP1 protein in the condition media, detected by RT-qPCR and ELISA, respectively. Each data point represents one donor-derived primary cell line for both NHLF ( n = 5 donors) and DHLF ( n = 5 donors), with each experiment performed in triplicates and represented as mean ± SD. Statistical analysis was performed using paired t -test and significance denoted as * p < 0.5 or ** p < 0.01 versus healthy NHLF, as shown.
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    <t>WISP1</t> is endogenously upregulated in primary human DHLF. WISP1 transcript and secreted protein are expressed in NHLF and DHLF. DHLF exhibit significantly higher ( A ) WISP1 mRNA transcript and ( B ) secreted WISP1 protein in the condition media, detected by RT-qPCR and ELISA, respectively. Each data point represents one donor-derived primary cell line for both NHLF ( n = 5 donors) and DHLF ( n = 5 donors), with each experiment performed in triplicates and represented as mean ± SD. Statistical analysis was performed using paired t -test and significance denoted as * p < 0.5 or ** p < 0.01 versus healthy NHLF, as shown.
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    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, <t>WISP1,</t> 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 μm (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 μm 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 ± 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.
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    BioVendor Instruments human wisp1 elisa kit
    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, <t>WISP1,</t> 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 μm (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 μm 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 ± 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.
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    Boster Bio human wisp1 ccn4 picokine tm elisa kit
    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, <t>WISP1,</t> 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 μm (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 μm 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 ± 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.
    Human Wisp1 Ccn4 Picokine Tm Elisa Kit, supplied by Boster Bio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    WISP1 is endogenously upregulated in primary human DHLF. WISP1 transcript and secreted protein are expressed in NHLF and DHLF. DHLF exhibit significantly higher ( A ) WISP1 mRNA transcript and ( B ) secreted WISP1 protein in the condition media, detected by RT-qPCR and ELISA, respectively. Each data point represents one donor-derived primary cell line for both NHLF ( n = 5 donors) and DHLF ( n = 5 donors), with each experiment performed in triplicates and represented as mean ± SD. Statistical analysis was performed using paired t -test and significance denoted as * p < 0.5 or ** p < 0.01 versus healthy NHLF, as shown.

    Journal: Cells

    Article Title: Stage-Dependent Fibrotic Gene Profiling of WISP1-Mediated Fibrogenesis in Human Fibroblasts

    doi: 10.3390/cells13232005

    Figure Lengend Snippet: WISP1 is endogenously upregulated in primary human DHLF. WISP1 transcript and secreted protein are expressed in NHLF and DHLF. DHLF exhibit significantly higher ( A ) WISP1 mRNA transcript and ( B ) secreted WISP1 protein in the condition media, detected by RT-qPCR and ELISA, respectively. Each data point represents one donor-derived primary cell line for both NHLF ( n = 5 donors) and DHLF ( n = 5 donors), with each experiment performed in triplicates and represented as mean ± SD. Statistical analysis was performed using paired t -test and significance denoted as * p < 0.5 or ** p < 0.01 versus healthy NHLF, as shown.

    Article Snippet: WISP1 (DY1627), IL-6 (D6050), and CCL-2 (DCP00) protein content was assessed using an enzyme-linked immunosorbent assay (ELISA) kit from R&D systems as per the manufacturer’s recommendations.

    Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Derivative Assay

    TGFβ induces WISP1 expression in primary lung and dermal fibroblasts. Stimulation with TGFβ (1 ng/mL) for 24 h significantly increases COL1A1, COL3A1, IL6, ACTA2, FN1, and WISP1 mRNA as compared to control in ( A ) NHLF, ( B ) DHLF, and ( C ) NHDF for 3 respective donors ( n = 3/each) analyzed by RT-qPCR. Notably, TGFβ decreased CCL2 in NHLF and DHLF, except in NHDF. Each experiment was independently performed in triplicates with three biological replicates ( n = 3) for each donor and represented as mean ± SD. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc analysis vs. vehicle control for respective donors. * denotes p < 0.5, ** denotes p < 0.01, *** denotes p < 0.001, **** denotes p < 0.0001 versus the unstimulated control, as shown.

    Journal: Cells

    Article Title: Stage-Dependent Fibrotic Gene Profiling of WISP1-Mediated Fibrogenesis in Human Fibroblasts

    doi: 10.3390/cells13232005

    Figure Lengend Snippet: TGFβ induces WISP1 expression in primary lung and dermal fibroblasts. Stimulation with TGFβ (1 ng/mL) for 24 h significantly increases COL1A1, COL3A1, IL6, ACTA2, FN1, and WISP1 mRNA as compared to control in ( A ) NHLF, ( B ) DHLF, and ( C ) NHDF for 3 respective donors ( n = 3/each) analyzed by RT-qPCR. Notably, TGFβ decreased CCL2 in NHLF and DHLF, except in NHDF. Each experiment was independently performed in triplicates with three biological replicates ( n = 3) for each donor and represented as mean ± SD. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc analysis vs. vehicle control for respective donors. * denotes p < 0.5, ** denotes p < 0.01, *** denotes p < 0.001, **** denotes p < 0.0001 versus the unstimulated control, as shown.

    Article Snippet: WISP1 (DY1627), IL-6 (D6050), and CCL-2 (DCP00) protein content was assessed using an enzyme-linked immunosorbent assay (ELISA) kit from R&D systems as per the manufacturer’s recommendations.

    Techniques: Expressing, Control, Quantitative RT-PCR

    WISP1 promotes a striking increase in primary human dermal fibroblast cell proliferation. ( A ) Treatment with WISP1 (1000 nM) shows a striking increase in cell proliferation ( p < 0.0001 via one-way ANOVA with Tukey’s post hoc analysis) detected by increased pRb positive cells as compared to control in NHDF. Representative 10× images were captured with the top image representing Hoechst nuclear stain, while the bottom panel represents pRb green fluorescence. PDGF-BB (100 nM) was used as an internal positive control to ensure assay reliability. The PDGF-BB dose response curve is shown in the . ( B ) % pRb positive cells were calculated by dividing pRb positive cells over total number of cells calculated by nuclear Hoechst staining. Three independent experiments ( n = 3) were performed in triplicates and represented as mean ± SD. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc analysis versus the vehicle-control condition and significance denoted as * p < 0.5, **** p < 0.0001.

    Journal: Cells

    Article Title: Stage-Dependent Fibrotic Gene Profiling of WISP1-Mediated Fibrogenesis in Human Fibroblasts

    doi: 10.3390/cells13232005

    Figure Lengend Snippet: WISP1 promotes a striking increase in primary human dermal fibroblast cell proliferation. ( A ) Treatment with WISP1 (1000 nM) shows a striking increase in cell proliferation ( p < 0.0001 via one-way ANOVA with Tukey’s post hoc analysis) detected by increased pRb positive cells as compared to control in NHDF. Representative 10× images were captured with the top image representing Hoechst nuclear stain, while the bottom panel represents pRb green fluorescence. PDGF-BB (100 nM) was used as an internal positive control to ensure assay reliability. The PDGF-BB dose response curve is shown in the . ( B ) % pRb positive cells were calculated by dividing pRb positive cells over total number of cells calculated by nuclear Hoechst staining. Three independent experiments ( n = 3) were performed in triplicates and represented as mean ± SD. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc analysis versus the vehicle-control condition and significance denoted as * p < 0.5, **** p < 0.0001.

    Article Snippet: WISP1 (DY1627), IL-6 (D6050), and CCL-2 (DCP00) protein content was assessed using an enzyme-linked immunosorbent assay (ELISA) kit from R&D systems as per the manufacturer’s recommendations.

    Techniques: Control, Staining, Fluorescence, Positive Control

    WISP1 increases CCL2 and IL6 in primary fibroblasts. Stimulation with WISP1 (1000 nM) for 24 h significantly increases CCL2 and IL6 gene expression in ( A ) NHLF, ( B ) DHLF, and ( C ) NHDF as compared to control. TGFβ (1 ng/mL) for 24 h served as a positive control and significantly increases COL1A1, COL3A1, IL6, ACTA2, FN1, and WISP1 versus vehicle control. ( D ) Stimulation with increasing WISP1 concentrations for 24 h initiates concentration-dependent increase in CCL2 and IL6 protein levels, detected via ELISA in the cell-supernatant of NHDF. Each experiment performed in duplicates with three biological replicates ( n = 3) and represented as mean ± SD. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc analysis vs. vehicle control for respective donors. * denotes p < 0.5, ** denotes p < 0.01, *** denotes p < 0.001, **** denotes p < 0.0001 versus the vehicle control, as shown.

    Journal: Cells

    Article Title: Stage-Dependent Fibrotic Gene Profiling of WISP1-Mediated Fibrogenesis in Human Fibroblasts

    doi: 10.3390/cells13232005

    Figure Lengend Snippet: WISP1 increases CCL2 and IL6 in primary fibroblasts. Stimulation with WISP1 (1000 nM) for 24 h significantly increases CCL2 and IL6 gene expression in ( A ) NHLF, ( B ) DHLF, and ( C ) NHDF as compared to control. TGFβ (1 ng/mL) for 24 h served as a positive control and significantly increases COL1A1, COL3A1, IL6, ACTA2, FN1, and WISP1 versus vehicle control. ( D ) Stimulation with increasing WISP1 concentrations for 24 h initiates concentration-dependent increase in CCL2 and IL6 protein levels, detected via ELISA in the cell-supernatant of NHDF. Each experiment performed in duplicates with three biological replicates ( n = 3) and represented as mean ± SD. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc analysis vs. vehicle control for respective donors. * denotes p < 0.5, ** denotes p < 0.01, *** denotes p < 0.001, **** denotes p < 0.0001 versus the vehicle control, as shown.

    Article Snippet: WISP1 (DY1627), IL-6 (D6050), and CCL-2 (DCP00) protein content was assessed using an enzyme-linked immunosorbent assay (ELISA) kit from R&D systems as per the manufacturer’s recommendations.

    Techniques: Gene Expression, Control, Positive Control, Concentration Assay, Enzyme-linked Immunosorbent Assay

    Recombinant WISP1 synergistically induces inflammatory markers along with TGFβ. ( A ) Primary human fibroblasts were stimulated with TGFβ (1 ng/mL) for 24 h for initiation, followed by WISP1 (1000 nM) for another 24 h as per the experimental layout. CCL2 and IL6 mRNA expression was significantly increased in ( B ) non-diseased lung and ( D ) dermal fibroblasts, but not in ( C ) IPF-diseased lung fibroblasts. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc analysis vs. vehicle control for respective donors. * denotes p < 0.5, ** denotes p < 0.01, *** denotes p < 0.001, **** denotes p < 0.0001 versus the unstimulated control, for three biological replicates ( n = 3) and represented as mean ± SD.

    Journal: Cells

    Article Title: Stage-Dependent Fibrotic Gene Profiling of WISP1-Mediated Fibrogenesis in Human Fibroblasts

    doi: 10.3390/cells13232005

    Figure Lengend Snippet: Recombinant WISP1 synergistically induces inflammatory markers along with TGFβ. ( A ) Primary human fibroblasts were stimulated with TGFβ (1 ng/mL) for 24 h for initiation, followed by WISP1 (1000 nM) for another 24 h as per the experimental layout. CCL2 and IL6 mRNA expression was significantly increased in ( B ) non-diseased lung and ( D ) dermal fibroblasts, but not in ( C ) IPF-diseased lung fibroblasts. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc analysis vs. vehicle control for respective donors. * denotes p < 0.5, ** denotes p < 0.01, *** denotes p < 0.001, **** denotes p < 0.0001 versus the unstimulated control, for three biological replicates ( n = 3) and represented as mean ± SD.

    Article Snippet: WISP1 (DY1627), IL-6 (D6050), and CCL-2 (DCP00) protein content was assessed using an enzyme-linked immunosorbent assay (ELISA) kit from R&D systems as per the manufacturer’s recommendations.

    Techniques: Recombinant, Expressing, Control

    WISP1 fibrotic gene signature in dermal fibroblasts. The volcano plot illustrates fibroblast genes upregulated (red) and downregulated (blue) upon stimulation with either ( A ) WISP1 alone or ( B ) in conjugation with TGFβ as compared to vehicle control (PBS) in NHDF ( n = 3). The plots representing statistically significant gene with |FC| > 1.5 and adj. p -value < 0.1 are plotted with the x-axis representing log2 fold change (FC) versus the y-axis −log10 p -value. ( C ) The heatmap represents pathways significantly associated with the corresponding treatment condition as percent of genes influenced and the list of genes used in the pathway analysis is highlighted in . ( D ) Scatterplot showing log2 fold change for the differentially expressed genes when comparing WISP1 + TGFβ treatment with WISP1 (x-axis) versus TGFβ (y-axis). ( E ) Venn diagram representing unique and overlapping set of genes between the WISP1, TGFβ, and WISP1 + TGFβ treatment paradigms. A comprehensive lists of genes is provided in the .

    Journal: Cells

    Article Title: Stage-Dependent Fibrotic Gene Profiling of WISP1-Mediated Fibrogenesis in Human Fibroblasts

    doi: 10.3390/cells13232005

    Figure Lengend Snippet: WISP1 fibrotic gene signature in dermal fibroblasts. The volcano plot illustrates fibroblast genes upregulated (red) and downregulated (blue) upon stimulation with either ( A ) WISP1 alone or ( B ) in conjugation with TGFβ as compared to vehicle control (PBS) in NHDF ( n = 3). The plots representing statistically significant gene with |FC| > 1.5 and adj. p -value < 0.1 are plotted with the x-axis representing log2 fold change (FC) versus the y-axis −log10 p -value. ( C ) The heatmap represents pathways significantly associated with the corresponding treatment condition as percent of genes influenced and the list of genes used in the pathway analysis is highlighted in . ( D ) Scatterplot showing log2 fold change for the differentially expressed genes when comparing WISP1 + TGFβ treatment with WISP1 (x-axis) versus TGFβ (y-axis). ( E ) Venn diagram representing unique and overlapping set of genes between the WISP1, TGFβ, and WISP1 + TGFβ treatment paradigms. A comprehensive lists of genes is provided in the .

    Article Snippet: WISP1 (DY1627), IL-6 (D6050), and CCL-2 (DCP00) protein content was assessed using an enzyme-linked immunosorbent assay (ELISA) kit from R&D systems as per the manufacturer’s recommendations.

    Techniques: Conjugation Assay, Control

    WISP1 fibrotic gene signature in lung fibroblasts. The volcano plot illustrates fibroblast genes upregulated (red) and downregulated (blue) upon stimulation with either ( A ) WISP1 alone or ( B ) in conjugation with TGFβ as compared to vehicle control (PBS) in NHLF ( n = 3). The plots representing statistically significant gene with |FC| > 1.5 and adj p -value < 0.1 are plotted with the x-axis representing log2 fold change (FC) versus the y-axis −log10 p -value. ( C ) The heatmap represents pathways significantly associated with the corresponding treatment condition as percent of genes influenced and the list of genes used in the pathway analysis is highlighted in . ( D ) Scatterplot showing log2 fold change for the differentially expressed genes when comparing WISP1 + TGFβ treatment with WISP1 (x-axis) versus TGFβ (y-axis). ( E ) Venn diagram representing unique and overlapping set of genes between the WISP1, TGFβ, and WISP1 + TGFβ treatment paradigms. A comprehensive lists of genes is provided in the .

    Journal: Cells

    Article Title: Stage-Dependent Fibrotic Gene Profiling of WISP1-Mediated Fibrogenesis in Human Fibroblasts

    doi: 10.3390/cells13232005

    Figure Lengend Snippet: WISP1 fibrotic gene signature in lung fibroblasts. The volcano plot illustrates fibroblast genes upregulated (red) and downregulated (blue) upon stimulation with either ( A ) WISP1 alone or ( B ) in conjugation with TGFβ as compared to vehicle control (PBS) in NHLF ( n = 3). The plots representing statistically significant gene with |FC| > 1.5 and adj p -value < 0.1 are plotted with the x-axis representing log2 fold change (FC) versus the y-axis −log10 p -value. ( C ) The heatmap represents pathways significantly associated with the corresponding treatment condition as percent of genes influenced and the list of genes used in the pathway analysis is highlighted in . ( D ) Scatterplot showing log2 fold change for the differentially expressed genes when comparing WISP1 + TGFβ treatment with WISP1 (x-axis) versus TGFβ (y-axis). ( E ) Venn diagram representing unique and overlapping set of genes between the WISP1, TGFβ, and WISP1 + TGFβ treatment paradigms. A comprehensive lists of genes is provided in the .

    Article Snippet: WISP1 (DY1627), IL-6 (D6050), and CCL-2 (DCP00) protein content was assessed using an enzyme-linked immunosorbent assay (ELISA) kit from R&D systems as per the manufacturer’s recommendations.

    Techniques: Conjugation Assay, Control

    WISP1 fibrotic gene signature in IPF-diseased fibroblast. The volcano plot illustrates fibroblast genes upregulated (red) and downregulated (blue) upon stimulation with either ( A ) WISP1 alone or ( B ) in conjugation with TGFβ as compared to vehicle control (PBS) in DHLF ( n = 3). The plots representing statistically significant gene with |FC| > 1.5 and adj p -value < 0.1 are plotted with the x-axis representing log2 fold change (FC) versus the y-axis −log10 p -value. ( C ) The heatmap represents pathways significantly associated with the corresponding treatment condition as percent of genes influenced and the list of genes used in the pathway analysis is highlighted in . ( D ) Scatterplot showing log2 fold change for the differentially expressed genes when comparing WISP1 + TGFβ treatment with WISP1 (x-axis) versus TGFβ (y-axis). ( E ) Venn diagram represents unique and overlapping set of genes between the WISP1, TGFβ, and WISP1 + TGFβ treatment paradigms. A comprehensive list of genes is provided in the .

    Journal: Cells

    Article Title: Stage-Dependent Fibrotic Gene Profiling of WISP1-Mediated Fibrogenesis in Human Fibroblasts

    doi: 10.3390/cells13232005

    Figure Lengend Snippet: WISP1 fibrotic gene signature in IPF-diseased fibroblast. The volcano plot illustrates fibroblast genes upregulated (red) and downregulated (blue) upon stimulation with either ( A ) WISP1 alone or ( B ) in conjugation with TGFβ as compared to vehicle control (PBS) in DHLF ( n = 3). The plots representing statistically significant gene with |FC| > 1.5 and adj p -value < 0.1 are plotted with the x-axis representing log2 fold change (FC) versus the y-axis −log10 p -value. ( C ) The heatmap represents pathways significantly associated with the corresponding treatment condition as percent of genes influenced and the list of genes used in the pathway analysis is highlighted in . ( D ) Scatterplot showing log2 fold change for the differentially expressed genes when comparing WISP1 + TGFβ treatment with WISP1 (x-axis) versus TGFβ (y-axis). ( E ) Venn diagram represents unique and overlapping set of genes between the WISP1, TGFβ, and WISP1 + TGFβ treatment paradigms. A comprehensive list of genes is provided in the .

    Article Snippet: WISP1 (DY1627), IL-6 (D6050), and CCL-2 (DCP00) protein content was assessed using an enzyme-linked immunosorbent assay (ELISA) kit from R&D systems as per the manufacturer’s recommendations.

    Techniques: Conjugation Assay, Control

    Schematic summary of WISP1-TGFβ crosstalk in regulation of fibrosis progression.

    Journal: Cells

    Article Title: Stage-Dependent Fibrotic Gene Profiling of WISP1-Mediated Fibrogenesis in Human Fibroblasts

    doi: 10.3390/cells13232005

    Figure Lengend Snippet: Schematic summary of WISP1-TGFβ crosstalk in regulation of fibrosis progression.

    Article Snippet: WISP1 (DY1627), IL-6 (D6050), and CCL-2 (DCP00) protein content was assessed using an enzyme-linked immunosorbent assay (ELISA) kit from R&D systems as per the manufacturer’s recommendations.

    Techniques:

    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 μm (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 μm 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 ± 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: 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 μm (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 μm 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 ± 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: human WISP1 ELISA kit , Elabscience , Cat#E-EL-H5542.

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

    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 μm. 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 μm 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 ± SEM, based on three biological replicates. Significance levels are denoted as ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.005.

    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: 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 μm. 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 μm 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 ± SEM, based on three biological replicates. Significance levels are denoted as ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.005.

    Article Snippet: human WISP1 ELISA kit , Elabscience , Cat#E-EL-H5542.

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

    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 ± SEM from three biological replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.005.

    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: 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 ± SEM from three biological replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.005.

    Article Snippet: human WISP1 ELISA kit , Elabscience , Cat#E-EL-H5542.

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

    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 μm. (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. (G) GSEAs of the TCGA PCa dataset revealed significant associations between high LIF and WISP1 expressions in prostate tissues with gene signatures representing metastatic prostate stroma-responsive signaling. NES, normalized enrichment score; FDR, false discovery rate. (H) A schematic summary of this study is presented. Crosstalk between PCa cells and prostate stromal cells within the tumor microenvironment (TME) may enhance the interaction between the CXCL5/CXCR2 and LIF/LIFR pathways. This interaction can potentially upregulate the secretion of LIF and WISP1 proteins, facilitated by the critical roles of STAT3 signaling-driven activation of the WISP1 and LIF genes.

    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: 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 μm. (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. (G) GSEAs of the TCGA PCa dataset revealed significant associations between high LIF and WISP1 expressions in prostate tissues with gene signatures representing metastatic prostate stroma-responsive signaling. NES, normalized enrichment score; FDR, false discovery rate. (H) A schematic summary of this study is presented. Crosstalk between PCa cells and prostate stromal cells within the tumor microenvironment (TME) may enhance the interaction between the CXCL5/CXCR2 and LIF/LIFR pathways. This interaction can potentially upregulate the secretion of LIF and WISP1 proteins, facilitated by the critical roles of STAT3 signaling-driven activation of the WISP1 and LIF genes.

    Article Snippet: human WISP1 ELISA kit , Elabscience , Cat#E-EL-H5542.

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

    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:

    Article Snippet: human WISP1 ELISA kit , Elabscience , Cat#E-EL-H5542.

    Techniques: Recombinant, Transfection, cDNA Synthesis, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, Software, ChIP-sequencing