primer and reverse transcriptase a primer pair specific and taq polymerase takara shiga japan cdna synthesis of 1 was used in for reaction of 20 u03bcl constituting 0 5 u taq dna polymerase 1 buffer and 1 mm dntp mix takara and specific primer pairs amplification by pcr was performed using a geneamp pcr system 2700 applied biosystems foster city calif usa under the following conditions 5 minutes at 94 u00b0 then 45 seconds at 94 u00b0 45 seconds at 56 u00b0 and 1 minute at 72 u00b0 and then subjected to 25 to 40 cycles final extension reaction was carried out at 72 u00b0 for 7 minutes table 1 shows the primer sequences used in the cdna amplification by for for primers were designed using primer3 program and were purchased from bioneer inc republic of korea daejeon pcr product was separated on a 1 5 agarose gel stained with ethidium bromide to ethidium bromide etbr visualized by gel doc 2000 uv trans illuminator bio rad laboratories hercules calif usa and then analyzed by using quantity one software bio rad laboratories each sample was tested at least three times and data representative was proposed cells were pre treated with 0 01 0 1 1 0 and 10 u03bcg ml of ec 18 respectively for 1 hour and antigen was processed and the incubated for 3 hours subsequently the rna was isolated after cell lysis cdna was prepared using reverse transcriptase reverse transcriptase with poly a primers on rna the design primers for il 4 was used in the pcr amplification table 1 gapdh was used.