5 15 dpp santa cruz sc 204305  (Santa Cruz Biotechnology)

Journal: Frontiers in Immunology

Article Title: Activated Platelets Convert CD14 + CD16 - Into CD14 + CD16 + Monocytes With Enhanced FcγR-Mediated Phagocytosis and Skewed M2 Polarization

doi: 10.3389/fimmu.2020.611133

Figure Lengend Snippet: Activated platelet-induced CD16 expression is mediated by phospho-SMAD3 and phospho-STAT3. (A) Immunoblot analysis of phosphorylated and total protein expression of STAT3 and SMAD3 in cell lysates from activated platelets alone, monocytes co-cultured with ADP-activated platelets, and monocytes alone at the indicated time points ( n = 3). (B) Representative contour plot of CD16 expression on monocytes co-cultured for 18 h with activated platelets in the presence of SIS3 (SMAD3 inhibitor) or 5,15-DPP (STAT3 inhibitor) (upper panel). Frequencies (%) of CD14 + CD16 + monocytes treated with SIS3 or 5,15-DPP were analyzed using flow cytometry (lower panel) ( n = 4). 0.1% DMSO was used as a vehicle. Bars show the mean ± S.E.M. * p < 0.05 and ** p < 0.01 by two-tailed paired t -test.

Article Snippet: In some experiments, CD14 + CD16 - monocytes were pre-incubate for 30 min at 37°C with 5,15-DPP (STAT3 inhibitor VIII), SIS3 (SMAD3 inhibitor), or SB431542 (TGF-βRI inhibitor) (all from MERCK, Burlington, MA) to examine the effect of TGF-β and IL-6-mediated signal transduction on CD16 induction by activated platelets.

Techniques: Expressing, Western Blot, Cell Culture, Flow Cytometry, Two Tailed Test

Journal: Archives of Toxicology

Article Title: Ethanol potentiates the genotoxicity of the food-derived mammary carcinogen PhIP in human estrogen receptor-positive mammary cells: mechanistic support for lifestyle factors (cooked red meat and ethanol) associated with mammary cancer

doi: 10.1007/s00204-018-2160-9

Figure Lengend Snippet: CYP2E1 mRNA expression mediated by PhIP in breast cells. CYP2E1 expression in MCF-7 and MDA-MB-231 cells ( a ) was measured by RT-qPCR. The involvement of miR378 and JAK/STAT3 pathway in MCF-7 cells treated with different concentrations of PhIP was assessed by qPCR ( b ). Data were normalized to expression of GAPDH and are shown relative to control 0.1% DMSO. Correlation of mRNA expression of CYP2E1 and STAT3 in PhIP-treated MCF-7 cells is shown in ( c ). Statistical significance was calculated by linear regression in GraphPad Prism 6, *** p < 0.001. Error bars represent SEM for independent cultures ( n = 3)

Article Snippet: For STAT3 inhibition, cells were co-treated for 24 h with PhIP and 25 μM STAT3 inhibitor (STAT3 inhibitor VIII 5, 15 diphenylporphyrin, Millipore, Feltham, UK).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Archives of Toxicology

Article Title: Ethanol potentiates the genotoxicity of the food-derived mammary carcinogen PhIP in human estrogen receptor-positive mammary cells: mechanistic support for lifestyle factors (cooked red meat and ethanol) associated with mammary cancer

doi: 10.1007/s00204-018-2160-9

Figure Lengend Snippet: Role of estrogen receptor (ER) and STAT3 in CYP2E1 induction by PhIP in MCF-7 cells: CYP2E1 mRNA expression in the presence of STAT3 inhibitor 25 µM ( a ), STAT3 mRNA expression in the presence of PhIP and ICI 182,780 in MCF7 cells ( b ), CYP2E1 expression in the presence of ER inhibitor ICI 182,780 (1 µM) ( c ). DMSO (0.2%) was used as vehicle control. Statistically significant differences between PhIP vs. PhIP & ICI 182, 780/STAT-3 inhibitor co-treated samples were assessed by Student’s t test in GraphPad Prism 6. *** p < 0.001, ** p < 0.01, * p < 0.05. Error bars represent the standard error of the mean (SEM) for independent cultures ( n = 3). Proposed mechanism of PhIP-mediated CYP2E1 induction ( d )

Article Snippet: For STAT3 inhibition, cells were co-treated for 24 h with PhIP and 25 μM STAT3 inhibitor (STAT3 inhibitor VIII 5, 15 diphenylporphyrin, Millipore, Feltham, UK).

Techniques: Expressing, Control

Journal: Archives of Toxicology

Article Title: Ethanol potentiates the genotoxicity of the food-derived mammary carcinogen PhIP in human estrogen receptor-positive mammary cells: mechanistic support for lifestyle factors (cooked red meat and ethanol) associated with mammary cancer

doi: 10.1007/s00204-018-2160-9

Figure Lengend Snippet: Proposed mechanism by which PhIP and ethanol induces genotoxicity in MCF-7 cells. PhIP can activate the AhR leading to upregulation of CYP1A2 /1B1 and increased metabolism of PhIP to genotoxic metabolites. PhIP can also upregulate CYP1B1 and CYP2E1 via ER-α through mir27b and STAT3, respectively. Additionally, ethanol can upregulate CYP2E1 expression. Collectively, these gene expression changes can lead to increased metabolic activation of carcinogenic substrates and higher levels of oxidative stress resulting in genotoxicity

Article Snippet: For STAT3 inhibition, cells were co-treated for 24 h with PhIP and 25 μM STAT3 inhibitor (STAT3 inhibitor VIII 5, 15 diphenylporphyrin, Millipore, Feltham, UK).

Techniques: Expressing, Gene Expression, Activation Assay