Page 91 - Helicobacter pylori and Gastric Cancer: From Tumor microenvironment to Immunotherapy

Noncanonical Hedgehog signaling
Noncanonical Hedgehog signalling
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FIGURE 2. Effects of Hedgehog stimulation on cellular signaling as determined by kinome profiling. Murine fibroblasts were stimulated with 1 μg/ml Shh. Subsequently cells were lysed and the resulting lysates were used to phosphorylate arrays of different kinase substrates employing 33P‐‐ATP and radioactivity incorporated in the different substrates was determined. Peptide substrates were allotted to elective signal transduction elements. The picture depicts the number of peptides significantly phosphorylated (which means the number of peptides that received a Markov “on” call - see experimental procedures) for each element. A darker color reflects more kinase activity towards substrate elements and the results reveal the effects of Hedgehog stimulation on cellular signal transduction, thus a black color means all peptides were significantly phosphorylated, whereas a white color means that no peptides allotted to this signal transduction in this experimental conditions were phosphorylated. Results were first statistically tested by a dichotomal analysis based on the number of Markov “on” calls observed in vehicle-and Shh-stimulated cultures. If statistically significant differences were noted the signal transduction category is highlighted with a red border and the level of significance observed is indicated in red. For signal transduction elements in which this very robust analysis fails to detect a statistically significant difference, a parametric test was performed. If this proved significant, the category is highlighted with an orange color and corresponding level of significance is depicted as well – in orange. The results provide a wealth of data on the effects of Hedgehog stimulation on cellular signaling.
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