Protein polarization underlies directional cell growth, cell morphogenesis, cell division, fate specification and differentiation in plant development. Analysis of in vivo protein dynamics reveals differential mobility of polarized proteins in plant cells, which may arise from lateral diffusion, local protein-protein interactions, and is restricted by protein-membrane-cell wall connections. The asymmetric protein dynamics may provide a mechanism for the regulation of asymmetric cell division and cell differentiation. In light of recent evidence for preprophase band (PPB)-independent mechanisms for orienting division planes, polarity proteins and their dynamics might provide regulation on the PPB at the cell cortex to directly influence phragmoplast positioning or alternatively, impinge on cytoplasmic microtubule-organizing centers (MTOCs) for spindle alignment. Differentiation of specialized cell types is often associated with the spatial regulation of cell wall architecture. Here we discuss the mechanisms of polarized signaling underlying regional cell wall biosynthesis, degradation, and modification during the differentiation of root endodermal cells and leaf epidermal guard cells.