Figure S1 - Enterocyte Proliferation and Signaling Are Constitutively Altered in Celiac Disease

EGF mRNA is increased in Fibroblasts from CD patients respect to controls. Quantitative PCR analysis of EGF mRNAs in fibroblasts from GFD CD patients and controls. RQ = relative quantity of mRNAs. Experiments were performed in triplicate and the data from 3 independent experiments were averaged. Columns represent the mean, and bars represent the standard deviation. * = pt-test). cDNAs were generated from total RNA of CD GFD and controls fibroblasts, using the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, USA) [25]. The resulting cDNA samples were subjected to 10 cycles of PCR amplification followed by real-time PCR using the TaqMan® PreAmp Master Mix Kit Protocol (Applied Biosystems, PN 4366127, Foster City, CA, USA). Each TaqMan Gene Expression assay consisted of two sequence-specific PCR primers and a TaqMan assay FAM-labelled MGB probe. Eighty ng of total cDNA (as total input RNA) was used for each replicate assay, and 3 replicates were run for each sample in a 96-well plate format. Beta-2-microglobulin (B2M) was used as the endogenous control gene. Assays were run with 2× Universal PCR Master Mix without UNG (uracil-N-glycosylase) on an Applied Biosystems 7300 Real-Time PCR System using universal cycling conditions (10 min at 95°C; 15 sec at 95°C, 1 min 60°C for 40 cycles). (DOCX)