Journal: Oncotarget

Article Title: Novel chemokine-like activities of histones in tumor metastasis

doi: 10.18632/oncotarget.11226

Figure Lengend Snippet: A-C. Histones (50 μg/ml, 24 hours) induced chemokine production and release as demonstrated with a Proteome Profiler™ Antibody Array in Hepa1-6 cells. D. Knockdown of NF-κB p65 and TLR4 (but not TLR2 and RAGE) in Hepa1-6 cells inhibited histone (50 μg/ml, 24 hours)-induced CCL9/10 release as demonstrated by ELISA assay (n=3, *, p<0.05 versus control shRNA group). E. Anti-CCL9/10 neutralizing antibody (1 mg/ml) partly inhibited histone (50 μg/ml, 24 hours)-induced Hepa1-6 cell migration (n=3, *, p<0.05).

Article Snippet: The production or release of chemokines was assayed using a Proteome ProfilerTM Antibody Chemokine Array Kit (#ARY020) from R&D Systems Inc. according to the manufacturer's instructions.

Techniques: Ab Array, Knockdown, Enzyme-linked Immunosorbent Assay, Control, shRNA, Migration

Journal: Oncotarget

Article Title: Novel chemokine-like activities of histones in tumor metastasis

doi: 10.18632/oncotarget.11226

Figure Lengend Snippet: A-B. Compared with the control group, TLR4 depletion (by using TLR4 −/− mice or TLR4 knockdown cells) or inhibition of histone release (by administration of 10 mg/kg heparin or 10 mg/kg H3 neutralizing antibody) limited the formation of lung metastasis (as shown in arrow) in mice based on tail vein injection of 3×10 6 Hepa1-6 cells (N=5 mice/group, *, p<0.05 versus control group). In contrast, control IgG (10 mg/kg) did not inhibit the formation of lung metastasis (B). C. Serum nucleosome levels were reduced after treatment with heparin in wildtype, but not in TLR4 −/− mice (N=5 mice/group, *, p<0.05 versus control group). D. Conceptual relationships between histone and tumor metastasis. Histone is a nuclear DAMP and can be released during cell injury or death. Once released, histone can promote cell migration and invasion through the TLR4-ERK-NF-κB pathway, which induces chemokine production and release.

Article Snippet: The production or release of chemokines was assayed using a Proteome ProfilerTM Antibody Chemokine Array Kit (#ARY020) from R&D Systems Inc. according to the manufacturer's instructions.

Techniques: Control, Knockdown, Inhibition, Injection, Migration

Journal: Cell Death & Disease

Article Title: Zebularine regulates early stages of mESC differentiation: effect on cardiac commitment

doi: 10.1038/cddis.2013.88

Figure Lengend Snippet: Characterization of HS-181 cell line after treatment with zebularine. ( a ) RT-PCR of cardiac markers after treatment with zebularine. Myh7, Myh6, Actc, cTnI and Serca2 present more expression after treatment. ( b ) Immunostaining of treated cells to detect cardiac-specific proteins. Scale bars; 50 μ m. ( c ) Blots of Proteome Profiler Array and the resulting quantification histograms demonstrating inhibition of pluripotency marker expression and ( d ) increased levels of mesodermic proteins after zebularine treatment (black arrows)

Article Snippet: Protein expression profiles were assayed using the specific human pluripotent Stem Cell array kit ‘Proteome Profiler Array' (R&D Systems Europe, Abingdon, UK; ARY010) following the manufacturer's instructions.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Immunostaining, Inhibition, Marker

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Neddylation Inhibition on Inflammation-Induced MMP9 Gene Expression in Esophageal Squamous Cell Carcinoma

doi: 10.3390/ijms22041716

Figure Lengend Snippet: Effect of tumor necrosis factor-alpha (TNF-α) and MLN4924 on the signaling pathways mediating matrix metalloproteinase 9 (MMP9) gene expression in esophageal squamous cell carcinoma (ESCC) cells. ( A ) Proteome profiling of the nuclear factor kappa B (NFκB) pathway by antibody array analyses in the KYSE150 cells treated with MLN4924 and TNF-α. Protein lysates from the untreated controls and cells treated with MLN4924 or TNF-α alone or in combination were analyzed using a human NFκB array (R&D). ( B ) Bar graphs showing the phosphorylation ratio (upper) and the protein level ratio (bottom) calculated after the semi-quantitative analysis of selected proteins. Results are presented as means ± SEM from duplicates. * p < 0.001, ** p < 0.05 vs. controls. The complete array is shown in . ( C ) Western blot analysis showing time-dependent activation of inhibitor of nuclear factor kappa B-alpha (IκB-α), NFκB/p65 and c-Jun in the KYSE150 cells treated with TNF-α (30 ng/mL). ( D ) MLN4924-dependent changes in the activation of IκB-α, NFκB/p65 and c-Jun as well as increasing levels of cyclin dependent kinase inhibitor 1A (CDKN1a/p21) protein in the KYSE150 cells within 24 h. ( E ) A dose-dependent effect of MLN4924 on activation of NFκB/p65 and c-Jun signaling pathways in the KYSE150 cells treated with TNF-α (30 g/mL) for 24 h. The effect of the KYSE70 cells treatment with different concentrations (0.25, 0.5, 1.0, 2.5 and 5.0 µM) of MLN4924 for 24 and 48 h is shown in .

Article Snippet: To determine the relative levels of 41 total and 4 serine/tyrosine phosphorylated proteins involved in NFκB signal transduction, the Proteome Profiler Human NF-κB Pathway Array (R&D Systems, ARY029) was used.

Techniques: Protein-Protein interactions, Gene Expression, Ab Array, Phospho-proteomics, Western Blot, Activation Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel method for the establishment of autologous skin cell suspensions: characterisation of cellular sub-populations, epidermal stem cell content and wound response-enhancing biological properties

doi: 10.3389/fbioe.2024.1386896

Figure Lengend Snippet: Proteomic analysis of the ACS secretome. Semiquantitative analysis using proteome arrays and quantitative detection by ELISA assays of soluble mediators secreted by ACS-derived cell populations cultured in serum-free medium at the indicated time points. Supernatants from cultures established from ACS were collected and analysed as detailed in the Methods. (A) Relative intensity values of densitometric analysis of ACS-secreted soluble factors using an angiogenesis array on days 1, 3 and 6 after isolation. (B) Relative intensity values of similar analysis using a cytokine proteome array on the same time-points post-isolation. (C) Detection of Interleukin-1 alpha, HMGB1 and Hsp90α in the ACS secretome by analyte-specific ELISA. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; ns, non-significance.

Article Snippet: Detection of analytes released in vitro from collected conditioned medium (at the indicated time points following culture initiation) from ACS-derived cultures was performed using an angiogenesis array kit (#ARY007) and a cytokine array kit (#ARY005B) (both from R&D Systems, supplied by Bio-Techne, Abingdon, United Kingdom).

Techniques: Enzyme-linked Immunosorbent Assay, Derivative Assay, Cell Culture, Isolation

Journal: bioRxiv

Article Title: IL-6R blockade with tocilizumab disrupts pericyte– and tumor cell–driven IL-6/STAT3 signaling, enhancing docetaxel efficacy in ER+ breast cancer

doi: 10.64898/2026.01.29.702661

Figure Lengend Snippet: a Cytokine expression. Fold change of cytokine array data highlighting IL-6, CXCL1, CXCL5, ST2, and GDF-15 from HBVP conditioned media. a.u. denotes arbitrary units. Data is representative of n = 2 independent experiments with technical duplicates. Mean ± s.e.m. shown. b IL-6 (pg/mL) from HBVP conditioned media quantified by IL-6 ELISA. Data is representative of n = 2 independent experiments with technical duplicates. Mean ± s.e.m. shown. c IL6 transcript levels in reference to GAPDH. a.u. denotes arbitrary units. Data is representative of n = 4 independent experiments with technical duplicates. Mean ± s.e.m. shown. P values were calculated by using an unpaired two-tailed t-test for a, b, and c . * P = 0.0245 ( a ), * P = 0.0134 ( b ), and * P = 0.0431 ( c ). * P ≤ 0.05. HBVP = human brain vascular pericytes. DTX = docetaxel.

Article Snippet: The conditioned media were collected after centrifuging for 5 minutes at RT and 600 g. The relative concentrations of 105 cytokines were measured using the Proteome Profiler Human XL Cytokine Array Kit (R&D Systems, part of Bio-Techne #ARY022B).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Caffeic acid selectively eliminates teratogenic human-induced pluripotent stem cells via apoptotic cell death.

doi: 10.1016/j.phymed.2022.154144

Figure Lengend Snippet: Fig. 3. Increase of caspase activities by CAA treatment in iPSCs. (A) Apoptosis-related proteins were detected by Western blotting in CAA-treated iPSCs and hDFs. Relative band intensities were calculated after normalization to β-actin expression. (B) iPSCs and hDFs were treated with 50 and 100 μM CAA for 24 h, and the caspase-3, −8, and −9 activities were determined. Relative caspase activity compared with CAA-untreated iPSCs and hDFs was expressed as the means ± SD (n = 3). (C) iPSCs were pre-treated with or without 10 μM of caspase-3 inhibitor (z-DEVD), caspase-8 inhibitor (z-IETD), caspase-9 inhibitor (z-LEHD), and pan-caspase inhibitor (z-VAD) for 30 min and then treated with 50 and 100 μM CAA. DMSO was treated as vehicle control. After 24 h, cells were stained with crystal violet solution and relative cell viability compared with CAA-untreated control iPSCs were expressed as the means ± SD (n = 3). **p < 0.01 vs. CAA-untreated control. ##p < 0.01 vs. Inhibitor-untreated control. Scale bar = 100 μM.

Article Snippet: The expression profile of human apoptosis-related proteins in CAA-treated or -untreated iPSCs was analyzed using a Proteome Profiler Human Apoptosis Array Kit A. Kim et al. Phytomedicine 102 (2022) 154144 (ARY009, R&D System, Minneapolis, MN, USA) according to the manufacturer’s instruction.

Techniques: Western Blot, Expressing, Activity Assay, Control, Staining