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Vision transformers (ViTs) encoding an image as a sequence of patches bring new paradigms for semantic segmentation.We present an efficient framework of representation separation in local-patch level and global-region level for semantic segmentation with ViTs. It is targeted for the peculiar over-smoothness of ViTs in semantic segmentation, and therefore differs from current popular paradigms of context modeling and most existing related methods reinforcing the advantage of attention. We first deliver the decoupled two-pathway network in which another pathway enhances and passes down local-patch discrepancy complementary to global representations of transformers. We then propose the spatially adaptive separation module to obtain more separate deep representations and the discriminative cross-attention which yields more discriminative region representations through novel auxiliary supervisions. The proposed methods achieve some impressive results: 1) incorporated with large-scale plain ViTs, our methods achieve new state-of-the-art performances on five widely used benchmarks; 2) using masked pre-trained plain ViTs, we achieve 68.9% mIoU on Pascal Context, setting a new record; 3) pyramid ViTs integrated with the decoupled two-pathway network even surpass the well-designed high-resolution ViTs on Cityscapes; 4) the improved representations by our framework have favorable transferability in images with natural corruptions. The codes will be released publicly.