*2.11. Quantitative PCR*

Total RNA was extracted from cells using TRIzol Reagent (Life Technologies, Carlsbad, CA, USA) according to the manufacturer's instructions. RNA was converted in cDNA using High-Capacity RNA-to-cDNATM kit (Applied Biosystems, Foster City, CA, USA). The cDNA concentrations were measured with a NanoVue Plus spectrophotometer (GE Healthcare, Chicago, IL, USA) and 50 ng cDNA was used. Real-time quantitative PCR was performed using the Power SYBR Green PCR Master Mix in Step One Plus Real-Time PCR System (Applied Biosystems). Primers for IL-3 (sense 5--ACAATGGTTCTTGCCAGCTCTAC-3- antisense 5--AGGAGCGGGAGCAGCAT-3-), IL-4 (sense 5--CAGGGTGCTTCGCAAATTTTAC-3- anti-sense 5--ACCGAGAACCCCAGACTTGTT-3-), IL-13 (sense 5--GCTCTCGCTTCGCTTGGTGGTC-3- anti-sense 5-CATCCGAGGCCTTTTGGTTACAG-3-), and GAPDH (sense 5--TTCACCACCATGGAGAAGGC-3- antisense 5-- GGCATGGACTGTGGTCATGA-3-) were based on validated sequences from Primer Bank [21]. GAPDH mRNA was used to normalize

data, with fold change calculated by the comparative Ct method (ΔΔCT method), as previously described [22]. Results are shown as mean ± s.d. of three independent experiments.
