Journal:

Article Title: Shiga Toxins Induce, Superinduce, and Stabilize a Variety of C-X-C Chemokine mRNAs in Intestinal Epithelial Cells, Resulting in Increased Chemokine Expression

doi: 10.1128/IAI.69.10.6140-6147.2001

Figure Lengend Snippet: Effect of Stx1 on the accumulation of various chemokine mRNAs. HCT-8 cells were treated for 4 h with either Stx1 (1 μg/ml) or heat-inactivated Stx1 (controls; 1 μg/ml). Total cellular RNA was isolated, and then radiolabeled cDNA was prepared from total cellular RNA using reverse transcriptase and cytokine-specific primers. Paired expression arrays were probed with radiolabeled cDNA from either Stx1-treated cells or control cells. Paired arrays were washed and exposed to film. Autoradiographs were scanned using an Agfa II scanner. (A) Pertinent sections from autoradiographs. Intensity of signal corrected for background was determined for each labeled chemokine gene on both Stx1-treated and control arrays using QuantityOne software (Bio-Rad). Results (B and C) are expressed as ratios between expression of the chemokine (B) or housekeeping mRNA (C) of interest in Stx1-treated and control cells. Abbreviations: β-2 micro, beta 2-microglobulin; HLA-A, major histocompatibility complex class I lymphocyte antigen (HLA-A 0201); HPRT, hypoxanthine phosphoribosyltransferase; L19, ribosomal protein L19; Tfr, transferrin receptor. Asterisk, ratio that is ≥100.

Article Snippet: Intensity of signal corrected for background was determined for each labeled chemokine gene on both Stx1-treated and control arrays using QuantityOne software (Bio-Rad).

Techniques: Isolation, Expressing, Labeling, Software