Figure 7. GM-CSF expression and secretion after acute smoke exposure is only up-regulated in lung tissue and BAL of mainstream CS-exposed mice.
(A) mRNA expression levels of GM-CSF in comparison with housekeeping control HPRT-1 were determined via quantitative real-time PCR using cDNA synthesized from lung tissue homogenate. Primers used are listed in Table 1. Relative mRNA expression is given as 2−ΔCt (ΔCt=Cttarget−Ctreference), relative changes compared with control are 2−ΔΔCt values (ΔΔCt=ΔCttreated−ΔCtcontrol). (B) Concentrations of secreted GM-CSF in BAL after 3 days of CS exposure were determined using a magnetic bead-based multiplex assay. For this assay, BAL fluid was concentrated (10×) by ultrafiltration in centrifugal filter devices. Results are means±S.D., one-way ANOVA following Bonferroni post-hoc test. ***P<0.001. FA, filter air; n=12. ns, not significant.