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This would be in contrast to previous findings in which both insula subdivisions have been implicated in stimulus-dependent processing (Albanese et al., 2007, Raij et al., 2005?and?Singer et al., 2004). The posterior insula in particular is considered a key region for nociceptive processing (Oertel et al., 2011). It is conceivable that this seeming discrepancy is a result of HDAC inhibitor the way in which our anatomical masks were defined. As highlighted before, all regions of interest were defined on the basis of anatomical landmarks, not functional contrasts, and all voxels within a given area entered the respective multivariate analysis. Some regions, such as the posterior insula, are known to be somatotopically organized (Brooks et al., 2005), suggesting that predictions from the entire region might not faithfully reflect the impact of a particular somatotopically relevant subregion. Under this view, the surprisingly low accuracies in the posterior insula might be a consequence of the functional heterogeneity across anatomical subdivisions included in our anatomical mask. We tested this hypothesis in an additional post-hoc analysis in which we utilized results from a previous study investigating the somatotopic organization of the insula ( Brooks et al., 2005) to consider those portions of the LMTK2 posterior insula that had been implicated in the processing of somatosensory stimuli applied to the same site as in the present experiment (main effect of pain, N?=?14, thresholded at U0126 p?=?0.001). Contrary to our initial hypothesis, the prediction accuracy obtained in foot-specific portions of the (left) posterior insula was not significantly above chance during the stimulation period. As indicated above, activity patterns in several pain-related regions allow for the prediction of pain, both during anticipation and during stimulation. However, this does not necessarily imply that predictions become even more accurate when considering several brain regions simultaneously. To investigate whether this might be the case, we carried out two additional analyses in which we examined increasing spatial scales of encoding. In the first analysis, we focused on pain-related brain regions and tested whether prediction accuracies would benefit from increasing the search space (i.e., increasing the potential complexity of the model) from the most predictive single region to combinations of multiple regions (Fig.?4). In the anticipation period, the most predictive single ROI (i.e., the right PAG) yielded a prediction accuracy of 56% (p?