A2A adenosine receptors and C/EBP{beta} are crucially required for IL-10 production by macrophages exposed to Escherichia coli -- Csóka et al. 110 (7): 2685 Data Supplement -

Supplemental materials for: Csoka et al -- Blood

Blood, Vol. 110, Issue 7, 2685-2695, October 1, 2007

A_2A adenosine receptors and C/EBPß are crucially required for IL-10 production by macrophages exposed to Escherichia coli
Blood Csóka et al. 110: 2685

Supplemental materials for: Csoka et al

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Figure S1. Generation of A_2B receptor KO mice (JPG, 62.2 KB) -
(A) A_2B KO mice were generated by the insertion of an IRES-lacZ-Neo cassette into exon 1 of the wild type A_2B allele using standard homologous recombination techniques in embryonic stem cells. Genotyping was conducted on genomic DNA isolated from tails using endogenous A_2B primers 1: 5′-GTACGTGGCGCTGGAGCTGGTTATC-3′ and 2: 5′-GCTCTGTGTGAGCACCAGCACGAAG-3′ and the neomycin primer 3: 5′-GGGTGGGATTAGATAAATGCCTGCTCT-3′. A combination of primers 1 and 2 give an amblicon of 247 bps corresponding to the wild type A_2B allele, while primers 2 and 3 yield a 417 bp amplicon corresponding to the mutant A_2B allele. (B) Total cellular RNA was isolated from the large intestine and bladder of A_2B receptor wild type (+/+), heterozygous (+/−) and KO (−/−) mice and real time RT-PCR was used to quantify A_2B receptor transcripts. Methods for RNA isolation and primers and PCR conditions can be found in reference 73. Results demonstrated half normal A_2B receptor transcripts in heterozygous tissues and the absence of A_2B receptor transcripts in KO tissues, verifying the presence of a null A_2B allele in these animals.