ccl17 rabbit polyclonal antibody  (Proteintech)

Journal: HemaSphere

Article Title: NOTCH1 fusions in pediatric T‐cell lymphoblastic lymphoma: A high‐risk subgroup with CCL17 (TARC) levels as diagnostic biomarker

doi: 10.1002/hem3.117

Figure Lengend Snippet: CCL17 (TARC) in NOTCH1 ‐rearranged patients. (A) CCL17 levels in pg/mL per patient, showing highly elevated CCL17 in blood of NOTCH1 ‐rearranged T‐LBL patients but none of the other patients. 10,000 pg/mL is the maximum measurable CCL17 level with used assay. Orange line in (A–C) represents maximum normal CCL17 level (1300 pg/mL) based on what has been described in Hodgkin lymphoma. ⁹ Patients that had a relapse are indicated with an asterisk. Patient TLBL049 developed a therapy‐related acute myeloid leukemia (double asterisk). (B) For three NOTCH1 ‐rearranged patients, blood CCL17 levels could be determined for a time point of remission after diagnosis, revealing normalized CCL17 levels in all three cases. (C) For four NOTCH1 ‐rearranged patients blood CCL17 levels were determined at time point of relapse and remission after relapse (second remission), revealing increased levels in three relapses that again normalized in second remission. (D) Staining for TARC using anti‐CCL17 antibody for four NOTCH1 ‐rearranged patients showing that T‐LBL cells do not express high levels of CCL17 based on immunohistochemistry.

Article Snippet: Immunohistochemistry staining for CCL17 was performed on the BOND‐III fully automated staining system (Leica) using CCL17 rabbit polyclonal antibody (ProteinTech Group).

Techniques: Biomarker Discovery, Staining, Immunohistochemistry

Journal: The Journal of Clinical Investigation

Article Title: HIC1 deletion promotes breast cancer progression by activating tumor cell/fibroblast crosstalk

doi: 10.1172/JCI99974

Figure Lengend Snippet: (A) Upper panel: schematic showing the coculture of MDA-231-LM2 BrCa cells with primary NAF10, CXCL14-activated NAF10, or primary CAF10 cells in a Transwell apparatus (0.4 μm pore size) for 4 days. Lower panel: Boyden chamber assay of MDA-231-LM2 cells that were treated as above. (B) Upper panel: Human XL Cytokine Array Kits (R&D Systems) were used to measure the levels of 102 cytokines in the CM from diverse fibroblasts. Cytokines that were upregulated in the CM of CXCL14-activated NAF10 and CAF10 cells are indicated by colored boxes; they include CCL17 (red), IL-5 (green), and angiopoietin-2 (blue). Black frames indicate the positive controls, and the dashed boxes indicate the negative controls in each membrane. Lower panel: table showing the relative signal intensities of the 3 selected cytokines noted above. The signal intensities were quantified by densitometry using ImageJ software and normalized to the intensity of the internal positive controls. (C) Boyden chamber assay of MDA-231-LM2 cells plated with rhCCL17, rhIL-5, and rh angiopoietin-2 in the lower chambers at 100 ng/ml for 20 hours. (D) Boyden chamber assay of MDA-231-LM2 cells that were cocultured with primary NAF10, CXCL14-activated NAF10, or primary CAF10 cells in a Transwell apparatus for 4 days in the presence or absence of α-CCL17 (1 μg/ml) or an isotype-matched IgG control. (E) MCF7 or MDA-231-LM2 cells were treated with various concentrations (0–100 ng/ml) of rhCCL17 for 4 days, and lysates of the cells were analyzed by Western blot using antibodies against E-cadherin, N-cadherin, and GAPDH. (F) MCF7 or MDA-231-LM2 cells were treated with rhCCL17 at 100 ng/ml for the indicated times (0, 5, 15, 30, and 60 minutes). Cell lysates were analyzed by Western blot with antibodies against p-Akt (Ser 473), Akt, p-GSK-3β (Ser9), GSK-3β, and GAPDH. (G) Knockdown of CCR4 expression by siRNA-3 in MCF7 and MDA-231-LM2 cells in the presence or absence of rhCCL17 at 100 ng/ml for 4 days. Cell lysates were analyzed by Western blot with antibodies against E-cadherin, N-cadherin, p-Akt (Ser 473), Akt, and GAPDH. Data are shown as mean ± SEM. n = 3 independent experiments. ***P < 0.001, 1-way ANOVA followed by Bonferroni’s post hoc test.

Article Snippet: High-density TMA of human BrCa clinical samples (catalog BRC2281) were obtained from a cohort of 228 patients and constructed by Superbiotek Inc. IHC staining was performed with specific antibodies against HIC1 (catalog bs15485R; Bioss), CXCL14 (catalog ab46010; Abcam), CCL17 (catalog ab195044; Abcam), GPR85 (catalog ab140783; Abcam), and CCR4 (catalog ab1669; Abcam).

Techniques: Boyden Chamber Assay, Software, Western Blot, Expressing

Journal: The Journal of Clinical Investigation

Article Title: HIC1 deletion promotes breast cancer progression by activating tumor cell/fibroblast crosstalk

doi: 10.1172/JCI99974

Figure Lengend Snippet: (A) Schematic showing the lung metastasis model of BrCa xenografts. MDA-231-LM2 BrCa cells mixed with primary NAF8 or CAF8 cells at a ratio of 3:1 were implanted bilaterally into the mammary fat pads of BALB/c nude mice (n = 12 per group). When the xenografts became palpable 2 weeks later, control IgG or αCCL17 (1 μg/mouse) was then injected via the tail vein every 3 days for 30 days; the mice were then euthanized. (B) Representative bioluminescence imaging of the harvested lungs (left) and quantification of their bioluminescent signals (right, n = 6). Data are shown as mean ± SD. *P < 0.05, 1-way ANOVA followed by Bonferroni’s post hoc test. (C) Representative immunohistochemical staining for human cytokeratin of each group’s lung tissues and for N-cadherin and vimentin in orthotopic xenografts. Dot plots show the mean values for the percentage of human cytokeratin-, N-cadherin–, or vimentin-positive cells with statistical evaluation (n = 5). Data are shown as mean ± SEM. n = 3 independent experiments. **P < 0.01; ***P < 0.001, 1-way ANOVA followed by Bonferroni’s post hoc test. (D) Kaplan-Meier survival curves of BALB/c nude mice that received the indicated treatments. *P < 0.05, log-rank test. (E) Kaplan-Meier plots of distant metastasis-free survival of systemically untreated patients (n = 543) stratified by CCL17 expression. Data were obtained from the Kaplan-Meier plotter database. P = 0.04, log-rank test (28).

Article Snippet: High-density TMA of human BrCa clinical samples (catalog BRC2281) were obtained from a cohort of 228 patients and constructed by Superbiotek Inc. IHC staining was performed with specific antibodies against HIC1 (catalog bs15485R; Bioss), CXCL14 (catalog ab46010; Abcam), CCL17 (catalog ab195044; Abcam), GPR85 (catalog ab140783; Abcam), and CCR4 (catalog ab1669; Abcam).

Techniques: Injection, Imaging, Immunohistochemical staining, Staining, Expressing

Journal: The Journal of Clinical Investigation

Article Title: HIC1 deletion promotes breast cancer progression by activating tumor cell/fibroblast crosstalk

doi: 10.1172/JCI99974

Figure Lengend Snippet: (A) Percentages of low and high expression of epithelial HIC1 (upper), stromal CXCL14 (middle), and stromal CCL17 (bottom) in benign breast tissue and various BrCa subtypes are shown as a pie chart. P vs. benign, χ2 test. (B) Representative IHC images showing the correlation between epithelial HIC1 expression and stromal CXCL14/CCL17 expression in 228 benign and malignant breast tumor samples. Broken lines indicate the margins of the tumor. (C) Schematic model showing how HIC1-mediated crosstalk between cancer cells and mammary fibroblasts promotes BrCa progression. Conditional deletion of HIC1 in the mouse mammary gland may contribute to premalignant transformation at the early stage of breast tumor formation. Moreover, chemokine CXCL14 secreted by HIC1-deleted BrCa cells binds to its cognate receptor GPR85 on mammary fibroblasts in the microenvironment, thereby activating the fibroblasts through the ERK1/2, Akt, and neddylation pathways. The activated fibroblasts in turn promote BrCa progression through induction of EMT by activation of the CCL17/CCR4 chemokine axis.

Article Snippet: High-density TMA of human BrCa clinical samples (catalog BRC2281) were obtained from a cohort of 228 patients and constructed by Superbiotek Inc. IHC staining was performed with specific antibodies against HIC1 (catalog bs15485R; Bioss), CXCL14 (catalog ab46010; Abcam), CCL17 (catalog ab195044; Abcam), GPR85 (catalog ab140783; Abcam), and CCR4 (catalog ab1669; Abcam).

Techniques: Expressing, Transformation Assay, Activation Assay

Journal:

Article Title: Characterization of the StcE Protease Activity of Escherichia coli O157:H7

doi: 10.1128/JB.01806-05

Figure Lengend Snippet: TABLE 2.

Article Snippet: Proteins were boiled for 5 min in SDS-PAGE sample loading buffer and analyzed by SDS-PAGE and immunoblot analysis using rabbit anti-human C1-INH polyclonal antibody (1:2,000) and an HRP-conjugated secondary antibody (1:2,000; Bio-Rad).

Techniques: Activity Assay