The present disclosure relates to the field of display technology, and provides a display apparatus, a display substrate, and a method of preparing a display substrate. The display substrate includes a display area and a frame area surrounding the display area, and further includes: a drive circuit layer provided on a base substrate; an insulating material structure provided on the drive circuit layer, located in the display area and at least a portion of the frame area, and provided with a step structure at an edge thereof; a light-emitting layer provided on the insulating material structure; an encapsulation layer covering the light-emitting layer; a first barrier wall provided on the drive circuit layer, and surrounding the insulating material structure, where the edge is an edge of the insulating material structure close to the first barrier wall; and a conductive connection structure provided inside the insulating material structure and electrically connected with the drive circuit layer. The present disclosure can avoid circuit defects.