Fluticasone and salmeterol inhibit C3a-induced production of chemokines.

<p><b>A</b>, LAD2 human mast cells were treated with fluticasone (1 uM) or salmeterol (1 uM) or in combination for 20 hr and stimulated with C3a (100 ng/mL) for 3 hr and CCL2, MIP-1β/CCL4 and CCL5 (<b>A</b>, <b>B</b> and <b>C</b> respectively) expression was measured by quantitative RT-PCR (n = 6, * = p<0.01 compared to untreated control, unless otherwise indicated).</p

Salmeterol inhibits neurokinin receptor expression.

<p>LAD2 human mast cells were treated with fluticasone (1 uM) or salmeterol (1 uM) or combination for 20 hr, and C3aR (<b>A</b>), neurokinin receptor 1(TARC1; <b>B</b>), 2 (TARC2; <b>C</b>) and 3 (TARC3; <b>D</b>) expression was measured by flow cytometry. Data are expressed as mean fluorescence intensity (MFI; n = 5, * = p<0.05 compared to untreated control).</p

Fluticasone and salmeterol synergistically inhibit mast cell degranulation.

<p><b>A</b>, LAD2 human mast cells were treated with 0.1, 1, 10 and 100 nM fluticasone (<b>A</b>, <b>B</b>, <b>C</b> and <b>D</b> respectively) and/or salmeterol for 20 hr, then stimulated with SP (0.5 ug/mL) for 30 min and β-hexosaminidase release was measured (n = 5, * = p<0.01).</p

Fluticasone and salmeterol synergistically inhibit SP-mediated production of TNF and chemokines.

<p>LAD2 mast cells were treated with fluticasone (1 uM) or salmeterol (1 uM) or in combination for 20 hr and stimulated with SP (1 ug/mL) for 24 hr and TNF, CXCL8 and CCL2 (<b>A</b>, <b>B</b> and <b>C</b> respectively) production was measured by ELISA (n = 3, * = p<0.01). <b>D</b>, CD34⁺-derived human mast cells were treated with fluticasone (1 uM) or salmeterol (1 uM) or in combination for 20 hr and stimulated with SP (1 ug/mL) for 24 hr and TNF production was measured by ELISA. (<b>E</b> and <b>F</b>) LAD2 mast cells were treated with fluticasone (1 uM) or salmeterol (1 uM) or combination for 20 hr and stimulated with C3a (100 ng/mL) for 24 hr and CCL2 and MIP-1β/CCL4 production was measured by ELISA. (<b>G</b> and <b>H</b>) CD34⁺-derived human mast cells were treated with fluticasone (1 uM) or salmeterol (1 uM) or in combination for 20 hr and stimulated with c3a (100 ng/mL) for 24 hr and CCL2 and MIP-1β/CCL4 production was measured by ELISA. (<b>I</b> and <b>J</b>) LAD2 mast cells were treated with fluticasone (1 uM) or salmeterol (1 uM) or in combination for 20 hr and stimulated with IgE/anti IgE (1 ug/mL/100 ug/ml) for 24 hr and CCL2 and CCL1 production was measured by ELISA (n = 3, * = p<0.01).</p

Fluticasone and salmeterol had no effect on FcεRI- and SP-mediated production of cysteinyl leukotrienes.

<p><b>A</b>, LAD2 mast cells were stimulated with C3a (100 ng/mL), SP (1 ug/mL), IgE (0.5 ug/mL) or IgE/anti-IgE (100 ug/mL) for 4 hr and CysLT production was measured by ELISA. <b>B</b>, LAD2 mast cells were stimulated with SP (1 ug/mL) for the indicated times and CysLT production was measured by ELISA. <b>C</b>, LAD2 mast cells were sensitized with IgE (0.5 ug/mL) and stimulated with anti-IgE (100 ug/mL) for 4 hr and CysLT production was measured by ELISA. <b>D</b>, LAD2 mast cells were stimulated with SP for 16 hr and CysLT production was measured by ELISA. <b>E</b>, LAD2 mast cells were sensitized with IgE (0.5 ug/mL), treated with fluticasone (0.1 uM) or salmeterol (1 uM) for 20 hr, stimulated with anti-IgE (100 ug/mL) for 4 hr and CysLT production was measured by ELISA. <b>F</b>, LAD2 mast cells were treated with fluticasone (0.1 uM) or salmeterol (1 uM) for 20 hr and stimulated with SP (1 ug/mL) for 16 hr and CysLT production was measured by ELISA. (n = 3, * = p<0.01).</p

Fluticasone and salmeterol inhibit FcεRI-mediated production of chemokines.

<p><b>A</b>, LAD2 human mast cells were sensitized with human myeloma IgE (0.5 ug/mL) for 16 hr, then treated with fluticasone (1 uM) or salmeterol (1 uM) or in combination for 20 hr and stimulated with anti-IgE (100 ug/mL) for 3 hr and CCL2 and CCL1 (<b>A</b> and <b>B</b> respectively) expression was measured by quantitative PCR (n = 6, * = p<0.01 compared to untreated control, unless otherwise indicated).</p

qPCR primer/probe sequences.

<p>qPCR primer/probe sequences.</p

Comparison of All Transcripts among the Control Groups.

<p>Pearson correlation and scatterplot matrix of log2 normalized expression of all transcripts in control groups, Con1-Con4B. The red solid line is the identity line. Again, the aberrant results of Con4A are due to less than optimal amplification prior to sequencing.</p

Expression of Ribosomal Subunits in the Control versus Amphetamine Groups.

<p>Pearson correlation and scatterplot matrix of log2 normalized expression of the ribosomal subunit proteins comparing each control group to its time-matched AMPH group. The red solid line is the identity line.</p

Citations for Gene Expression within Leukocyte Cell Types.

<p>Citations for Gene Expression within Leukocyte Cell Types.</p