insolution nf-kb activation inhibitor  (Millipore)

Journal: PLoS ONE

Article Title: The Gene Expression Response of Chronic Lymphocytic Leukemia Cells to IL-4 Is Specific, Depends on ZAP-70 Status and Is Differentially Affected by an NFκB Inhibitor

doi: 10.1371/journal.pone.0109533

Figure Lengend Snippet: (A) Scheme of the strategy used to identify the IL-4 targets in patients and in controls. (B) Heat maps for expression of IL-4 targets which had: top panel , similar changes in CLL and NBC and above 3-fold change for both; center panel , significantly higher changes in CLL and above 3-fold change in CLL; bottom panel , significantly higher changes in NBC and above 3-fold change in NBC. IL-4 targets are ordered alphabetically. In the event that several probes represent the same gene, only one is shown. Relative expression levels are depicted according to the shown log 2 color scale.

Article Snippet: In selected patients, additional fractions were treated with IL-4 plus InSolution NF-kB activation inhibitor [6-Amino-4-(4-phenoxyphenylethylamino)quinazoline] (Merck, Nottingham, UK) at 1 µM and 10 µM.

Techniques: Expressing

Journal: PLoS ONE

Article Title: The Gene Expression Response of Chronic Lymphocytic Leukemia Cells to IL-4 Is Specific, Depends on ZAP-70 Status and Is Differentially Affected by an NFκB Inhibitor

doi: 10.1371/journal.pone.0109533

Figure Lengend Snippet: Box whiskers representations of qPCR validations of 14 IL-4 targets representative of (A) restricted to CLL; (B) restricted to NBC; and (C) common to CLL and NBC. QPCR data are expressed as −ΔΔCt. IL-4 targets are ordered alphabetically.

Article Snippet: In selected patients, additional fractions were treated with IL-4 plus InSolution NF-kB activation inhibitor [6-Amino-4-(4-phenoxyphenylethylamino)quinazoline] (Merck, Nottingham, UK) at 1 µM and 10 µM.

Techniques:

Journal: PLoS ONE

Article Title: The Gene Expression Response of Chronic Lymphocytic Leukemia Cells to IL-4 Is Specific, Depends on ZAP-70 Status and Is Differentially Affected by an NFκB Inhibitor

doi: 10.1371/journal.pone.0109533

Figure Lengend Snippet: Triangular heat map representing the pairwise correlation coefficients (R) of the IL-4 upregulated targets between themselves. The IL-4 targets are ordered according to their correlation coefficients with ZAP-70, which are represented at the left and bottom sides. Cut-off values for positive or negative correlations with ZAP-70 were set at 0.4 and −0.4, respectively. In the event that several probes represent the same gene, only one is shown. Correlation coefficients are depicted according to the shown color scale. Inset shows Pearson correlation analysis between ZAP-70 levels by microarray (expressed as log 2 ratios) and by qPCR (expressed as –ΔΔCt ratios).

Article Snippet: In selected patients, additional fractions were treated with IL-4 plus InSolution NF-kB activation inhibitor [6-Amino-4-(4-phenoxyphenylethylamino)quinazoline] (Merck, Nottingham, UK) at 1 µM and 10 µM.

Techniques: Microarray

Journal: PLoS ONE

Article Title: The Gene Expression Response of Chronic Lymphocytic Leukemia Cells to IL-4 Is Specific, Depends on ZAP-70 Status and Is Differentially Affected by an NFκB Inhibitor

doi: 10.1371/journal.pone.0109533

Figure Lengend Snippet: (A) Apoptosis of NBC and CLL cells cultured for 18 h in the absence or presence of IL-4, and CLL cells cultured with IL-4 plus an NFκB activation inhibitor (NFκBi) at 1 µM and 10 µM. CLL are represented together and also separated in ZAP-70 positive and negative. Apoptosis was measured as the percentage of cells labelled with Annexin V ( top panel ). The bottom panel represents results in CLL patients after subtracting the percentage of apoptotic cells of the Ctrl samples to the IL-4, IL-4 plus 1 µM NFκBi, and IL-4 plus 10 µM NFκBi samples. T tests were used to compare the levels of apoptosis between cell types and conditions. When differences were significant the p-values are indicated as follows: *, p<0.05; **, p<0.01; ***, p<0.001. (B) Expression of the ZAP-70 Pos IL-4 targets SUSD2 and CCR2, and (C) of the ZAP-70 Neg IL-4 targets AUH, LY75, NFIL3, and BCL6), measured by qPCR, in 7 ZAP-70 positive and 8 ZAP-70 negative patients. The ratios for expression of IL-4 targets following treatment with IL-4 alone, IL-4 plus NFκBi at 1 µM, and IL-4 plus NFκBi at 10 µM, compared to Ctrl, are represented. P-values are depicted as in A.

Article Snippet: In selected patients, additional fractions were treated with IL-4 plus InSolution NF-kB activation inhibitor [6-Amino-4-(4-phenoxyphenylethylamino)quinazoline] (Merck, Nottingham, UK) at 1 µM and 10 µM.

Techniques: Cell Culture, Activation Assay, Expressing

Journal: PLoS ONE

Article Title: The Gene Expression Response of Chronic Lymphocytic Leukemia Cells to IL-4 Is Specific, Depends on ZAP-70 Status and Is Differentially Affected by an NFκB Inhibitor

doi: 10.1371/journal.pone.0109533

Figure Lengend Snippet: Adaptation of the pathway map entitled “Immune response_IL-4 – anti-apoptotic action” from MetaCore from Thomson Reuters. The map has been simplified leaving the minimal elements for activation of transcription factors essential in regulation of gene expression. ZAP-70 and its interactions with members of the pathway reported in the MetaCore database have been added to suggest its potential involvement in the pathway, and a link has been added for the reported interaction between NFκB and STAT6 . Red arrows indicate that IL4R, SOCS1, RPS6AK2, and NFKBIZ are components of the pathway identified as IL-4 upregulated targets in this study. Other genes of the pathway were significantly regulated by cell culture, such as AKT3 (up) and GRB2 (down).

Article Snippet: In selected patients, additional fractions were treated with IL-4 plus InSolution NF-kB activation inhibitor [6-Amino-4-(4-phenoxyphenylethylamino)quinazoline] (Merck, Nottingham, UK) at 1 µM and 10 µM.

Techniques: Activation Assay, Expressing, Cell Culture