A display substrate includes a substrate, a pixel-defining layer configured to define a plurality of sub-pixel regions, and a plurality of sub-pixels having at least two colors, each arranged within each of the plurality of sub-pixel regions. The pixel-defining layer and the plurality of sub-pixels are disposed over the substrate. The pixel-defining layer includes a plurality of first pixel-defining portions and a plurality of second pixel-defining portions. Each of the plurality of first pixel-defining portions is configured to separate neighboring sub-pixels of different colors. Each of the plurality of second pixel-defining portions is configured to separate neighboring sub-pixels of the same color. At least one of the plurality of second pixel-defining portions has a smaller width than any one of the plurality of first pixel-defining portions.