Journal: Oncotarget

Article Title: PTEN mediates the cross talk between breast and glial cells in brain metastases leading to rapid disease progression.

doi: 10.18632/oncotarget.14047

Figure Lengend Snippet: Figure 7: Effects of glial cell cocultures on cytokine secretion. Control and PTEN overexpressing MDA-MB-231 BR cells were seeded in transwell inserts and cocultured for 48 h with astrocytes (A) or microglia (B). Antibody-spotted membranes were incubated with conditioned media of these cultures and levels of secreted proteins were visualized by HRP-conjugated antibodies. Proteins with a more than 2-fold expression difference are shown by open boxes. Results were verified by qPCR analyses (C). Astrocyte coculture reduced EGF and GM-CSF expression significantly in PTEN overexpressing (PTEN CoA) compared to control cells (CTL CoA, C). Astrocytes express the GM- CSF receptor (CSFR2A) only when cocultured with tumor cells. Microglia coculture reduced VEGFA expression in PTEN overexpressing (PTEN CoMG) compared to control cells (CTL CoMG). Student’s t-test was used to analyze statistical significance. *p < 0.05.

Article Snippet: Cytokine secretion was estimated from CM by Proteome ProfilerTM Antibody Array (Human XL Cytokine Array Kit, ARY022, R&D Systems, Wiesbaden, Germany) according to manufacturer’s instruction.

Techniques: Control, Incubation, Expressing

Journal: British Journal of Cancer

Article Title: Antibody targeting of CD24 efficiently retards growth and influences cytokine milieu in experimental carcinomas

doi: 10.1038/bjc.2013.102

Figure Lengend Snippet: Effects of SWA11 mAb treatment on intratumoural cytokine milieu in A549 tumours. ( A ) The levels of 40 different cytokines were determined in tumour lysates using a cytokine protein arrays. The level of cytokines was quantified using ImageJ software. Representative arrays from two independent SWA11 mAb treatment experiments are shown. ( B ) Protein levels of CCL5/RANTES, CXCL9/MIG and CCL2/MCP-1 in tumour lysates ( n =5 per group) were determined by ELISA. Data are normalised to the total protein concentration in tumour lysates. * P <0.05.

Article Snippet: Relative levels of 40 different cytokines and chemokines in xenograft tumour lysates were evaluated using a mouse cytokine antibody array kit (Proteome Profiler, R&D Systems) according to the manufacturer's recommendations.

Techniques: Software, Enzyme-linked Immunosorbent Assay, Protein Concentration

Journal: British Journal of Cancer

Article Title: Antibody targeting of CD24 efficiently retards growth and influences cytokine milieu in experimental carcinomas

doi: 10.1038/bjc.2013.102

Figure Lengend Snippet: Effects of SWA11 mAb treatment on intratumoural cytokine milieu in SKOV3ip tumours. ( A ) The levels of 40 different cytokines were determined in tumour lysates using cytokine protein arrays. The level of cytokines was quantified using ImageJ software. Representative arrays from two independent SWA11 mAb treatment experiments are shown. ( B ) Protein levels of CCL5/RANTES, CXCL9/MIG and CCL2/MCP-1 in tumour lysates ( n =5–6 per group) were determined by ELISA. Data are normalised to the total protein concentration in tumour lysates.

Article Snippet: Relative levels of 40 different cytokines and chemokines in xenograft tumour lysates were evaluated using a mouse cytokine antibody array kit (Proteome Profiler, R&D Systems) according to the manufacturer's recommendations.

Techniques: Software, Enzyme-linked Immunosorbent Assay, Protein Concentration

Journal: British Journal of Cancer

Article Title: Antibody targeting of CD24 efficiently retards growth and influences cytokine milieu in experimental carcinomas

doi: 10.1038/bjc.2013.102

Figure Lengend Snippet: Effects of combined SWA11 mAb and gemcitabine treatment on A549 tumour growth and intratumoural cytokine milieu. ( A ) SCID beige mice with established xenograft A549 lung carcinomas received treatment with SWA11 mAb (10 mg kg −1 ) or IgG 2A (10 mg kg −1 ) followed by gemcitabine at a dose of 12 mg kg −1 (10% of maximal tolerated dose) 1 day later ( n =5 per group). Control animals received SWA11 mAb (10 mg kg −1 ), IgG 2A (10 mg kg −1 ) or gemcitabine (12 mg kg −1 ) alone ( n =5 per group). Treatment was repeated three times with a 5-day interval. External size of A549 tumours was measured with a caliper. * P <0.05. Note that part of the results from this experiment are also shown in for illustration. ( B ) Effects of SWA11 mAb and gemcitabine treatment on the levels of 40 different cytokines were determined in A549 tumour lysates using a cytokine protein arrays. The level of cytokines was quantified using ImageJ software.

Article Snippet: Relative levels of 40 different cytokines and chemokines in xenograft tumour lysates were evaluated using a mouse cytokine antibody array kit (Proteome Profiler, R&D Systems) according to the manufacturer's recommendations.

Techniques: Control, Software

Journal: Communications Biology

Article Title: Macrophage migration inhibitory factor is overproduced through EGR1 in TET2 low resting monocytes

doi: 10.1038/s42003-022-03057-w

Figure Lengend Snippet: a , b Cytokine profile arrays of supernatant collected from cord blood CD34 + cells infected with SCR - and TET2- shRNA-GFP lentiviruses, sorted on GFP expression, and induced to differentiate with stem cell factor (SCF), interleukin-3 (IL-3), Fms-related tyrosine kinase 3 ligand (FLT3L) and granulocyte-colony stimulating factor (G-CSF). a Representative cytokine array with supernatants collected at day 10 of differentiation. The rectangle points to MIF detection. b Quantification of MIF signals, normalized to positive controls. Data are mean +/− SEM of three independent experiments. Paired t test: * P < 0.05. c MIF concentrations determined by ELISA in the supernatant of cells induced to differentiate for indicated time. Data are mean +/− SEM of indicated independent experiments (day 5: n = 6; day 7: n = 5; day 8: n = 3; day 10: n = 7). Paired t test: * P < 0.05; *** P < 0.001. d RT-qPCR analysis of MIF mRNA expression in four TET2 -depleted ( TET2 shRNA, gray bars) and control ( SCR shRNA, black bars) human leukemic cell lines. Data are mean +/− SEM of three biological replicates. Unpaired t test: * P < 0.05; *** P < 0.001; **** P < 0.0001. e Immunoblot of SCR or TET2 shRNA infected leukemic cell lines sorted on GFP expression. Lower panels, quantification after actin normalization using Image J software. f MIF concentrations determined by ELISA in the supernatants of kasumi-1 ( n = 5), M07e ( n = 5), UT-7 ( n = 4) and TF-1 ( n = 5) cells transduced 24 h before with SCR (black squares) or TET2 (gray squares) shRNA. Data are mean +/− SEM of indicated biological replicates. Unpaired-t test: * P < 0.05; ** P < 0.01. g MIF concentrations determined by ELISA in the plasma of two Tet2 -deficient models (1–3 months): K427 knock-out model (wt/wt or Tet2 +/+ n = 4, wt/exc or Tet2 +/− n = 5 and exc/exc or Tet2 −/− n = 5); ANO knock-down model (wt/wt or Tet2 + /+ n = 5, wt/LacZ or Tet2 +/− n = 5 and LacZ/LacZ or Tet2 −/− n = 6). Data are mean +/− SEM of indicated biological replicates. Dunnett’s multiple comparison tests using wt/wt as control: * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: CD34 + collected supernatants were analyzed using human cytokine antibody array (panel A; R&D Systems).

Techniques: Infection, shRNA, Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Control, Western Blot, Software, Clinical Proteomics, Knock-Out, Knockdown, Comparison

Journal: Journal of the Chinese Medical Association : JCMA

Article Title: Identification of early response to hypertonic dextrose prolotherapy markers in knee osteoarthritis patients by an inflammation-related cytokine array

doi: 10.1097/JCMA.0000000000000693

Figure Lengend Snippet: Cytokine antibody array and hypertonic dextrose prolotherapy treatment of knee osteoarthritis. Human 274 cytokines were blotted onto a membrane and arrayed three times following the manufacturer’s protocol. The intensities of the relative expression levels of cytokines were quantified by densitometry (VersaDoc imaging system; Bio-Rad). The density value of each test sample was normalized to as described from Human Cytokine Antibody Array C6, C7, C8, C9, C10 (A) and graphed (B). Antibody arrays were used to examine the growth factors secreted. The arrays were scanned and quantified, and the levels were normalized to those of the positive controls.

Article Snippet: To explore this mechanism of paracrine regulation, the human chemokine and cytokine antibody array (Ray Biotech Inc., Norcross, GA, USA), including 274 specific antibodies, was used.

Techniques: Ab Array, Membrane, Expressing, Imaging