stat3 inhibitor cpd188  (Merck & Co)

Journal: Oncoimmunology

Article Title: Prostaglandin E 2 -mediated adenosinergic effects on CD14 + cells: Self-amplifying immunosuppression in cancer

doi: 10.1080/2162402X.2016.1268308

Figure Lengend Snippet: pSTAT3 inhibitor-mediated enhancement of CD73 induction and the function of CD73-expressing CD14+ cells. (A) Means + SD of pSTAT3 or isotype mfi values are shown from triplicate CD14+ cell samples at different times after sPE-treatment. (B) CD73 expression on CD14+ cells 48 h after treatment with sPE in the presence or absence of pSTAT3-inhibitor (STAT3i) Cpd188 (10 μM). Means + SD of CD73 mfi from triplicate samples are shown. (C) pSTAT3 (left panel) or pSTAT1 (right panel) or isotype control mfi values are shown 30 min after treatment with sPE in the presence or absence of Cpd188 (STAT3i) as described above. Means + SD of triplicates. (D) CD14+ healthy donor cells were treated with sPE or NECA (250 ng/mL) in the presence or absence of 10 ng/mL IL-6. Mfi of CD73 expression 48 h later is shown as mean + SD of triplicate samples. (E) CD14+ cells were treated with sPE overnight. LPS-induced TNFα production was measured by intracellular cytokine staining in the presence or absence of AMP (200 μM) or adenosine receptor A2A- and A2B-inhibitors (A2Ri; 200 nM each), as indicated under the graph. Means + SD of the mfi of TNFα from triplicate samples are shown. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Cpd188 (573125; STAT3-inhibitor) was obtained from Calbiochem and Anisomycin (A9789) from Sigma.

Techniques: Expressing, Staining

Journal: Oncoimmunology

Article Title: Prostaglandin E 2 -mediated adenosinergic effects on CD14 + cells: Self-amplifying immunosuppression in cancer

doi: 10.1080/2162402X.2016.1268308

Figure Lengend Snippet: The mechanism of CD73 induction on CD14+ cells. This cartoon summarizes the results of CD73-induction by adenosine, forskolin and PGE2, the inhibition of induction (dotted lines) by (1) adenosine receptor inhibitors, (2) PGE2 receptor inhibitors, (3) a PKA-inhibitor and (4) a p38-inhibitor. Negative regulation by pSTAT3 was demonstrated by a STAT3-inhibitor (5).

Article Snippet: Cpd188 (573125; STAT3-inhibitor) was obtained from Calbiochem and Anisomycin (A9789) from Sigma.

Techniques: Inhibition

Journal: Oncoimmunology

Article Title: Tumor stroma-derived factors skew monocyte to dendritic cell differentiation toward a suppressive CD14 + PD-L1 + phenotype in prostate cancer

doi: 10.4161/21624011.2014.955331

Figure Lengend Snippet: STAT3 and IL-6 blocking prevents altered DC differentiation caused by conditioned media from prostate cancer stroma. To determine the role of IL-6 and STAT3 in mediating responses to prostate cancer stroma conditioned media (PCaSt-CM), the phosphorylation status of STAT3 (A–B) and the effects of blockade of STAT3 and IL-6 signaling (C-E) were analyzed using monocytes treated with 50% PCaSt-CM plus GM-CSF/IL-4 with or without inhibitor, as indicated. (A and B) STAT3 (signal transducer and activator of transcription-3) phosphorylation (pSTAT3) was measured by flow cytometry. (i) Representative histograms showing pSTAT3 levels on the Y705 (A) or S727 residue (B). CD14+ cells were treated with GMCSF/IL-4 and exposed to 0% (DC) or 50% (sDC) conditioned medium from prostate cancer stromal cultures (PCaSt-CM) for 10 min. (ii) The mean ± SEM percentages of pSTAT3+ monocytes as in (i) after 0, 5, 10, 30 and 60 min (triplicates, sDC generated with 2 independent stromal conditioned media). (C) GM-CSF/IL-4 treatment downregulates CD14 expression on sDC in the presence of STAT3 Inhibitor IX Cpd188. (i) Representative dot plots without (left) and with (right) the STAT3 inhibitor. (ii) CD14 expression on day 3 DC (first bar) or sDC pre-treated with the STAT3 inhibitor (0, 0.5,1, 5, 10 μM) (mean + SEM; triplicates). (D) (i) Representative histogram; PD-L1 expression on day 5 DC or sDC (see above) treated with the STAT3 inhibitor (5μM Cpd188). (ii) Summary of the effect of the STAT3 inhibitor on PD-L1 expression. Bars represent the mean + SEM of PD-L1 expression on a single donor's DC or sDC, the latter treated with 50% PCaSt-CM from 3 different cultures (triplicates) in the presence (gray) or absence (black) of STAT3 inhibitor as in (i). (E). DC or sDC were cultured in the presence or absence of α-IL-6 and/or α-IL10 blocking antibodies (1 or 5 μg/mL) alone or together, for 5 d PD-L1 expression was assessed by flow cytometry. Mean + SEM from triplicate samples are shown. Statistical analysis was performed by two-way ANOVA; ns, not significant; ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05 .

Article Snippet: Monocytes were cultured for 3 d in SCBM+50% PCaSt-CM, containing 50 ng/mL GM-CSF and 500 U/mL IL-4, and in the presence of varying concentrations (0.5–10 μM) of STAT3 Inhibitor IX Cpd188 (Millipore), dissolved in dimethyl sulfoxide (DMSO).

Techniques: Blocking Assay, Phospho-proteomics, Flow Cytometry, Residue, Generated, Expressing, Cell Culture