Beginning in the First Intermediate Period of Egyptian history (c. 2181–2055 BC), Imsety, Hapy, Duamutef, and Qebehsenuef were especially connected with the four canopic jars that housed the internal organs that were removed from the body of the deceased during the process of mummification.

Most commonly, Imsety protected the liver, Hapy the lungs, Duamutef the stomach, and Qebehsenuef the intestines, but this pattern often varied.

Although they were increasingly closely associated with the internal organs, they continued to appear in burial equipment even after the use of canopic jars was abandoned in the Ptolemaic Period (303–30 BC), disappearing only in the fourth century AD with the extinction of the ancient Egyptian funerary tradition.

[2] In numerous sources, such as Spell 541 of the Pyramid Texts, they are stated to be the children of Horus, one of the major deities of the Egyptian pantheon.

In the tenth section of the New Kingdom Book of Gates, a funerary text that depicts the underworld in detail, the four sons are portrayed holding chains that bind the malign beings called "wmmtj-snakes".

Burials in which the jars and organs survive show that the most common arrangement was for Imsety to guard the liver, Hapy the lungs, Duamutef the stomach, and Qebehsenuef the intestines, but many variations are known.

[3] The sons of Horus themselves were thought to be under the protection of four goddesses, usually Isis for Imsety, Nephthys for Hapy, Neith for Duamutef, and Serqet for Qebehsenuef.

On some coffins, Imsety and Hapy remained at the head and Duamutef and Qebehsenuef at the foot, but in each pair, the deity who had been on the left side was moved to the right, and vice versa.

In other cases, each of the sons of Horus appeared on a side wall of the coffin or canopic chest: Imsety in the south, Hapy in the north, Duamutef in the east, and Qebehsenuef in the west.

[27] In the Eighteenth Dynasty (c. 1550–1292 BC), a few canopic jars were given varying heads: Imsety was portrayed as a man, Hapy as a baboon, Duamutef as a jackal and Qebehsenuef as a falcon.

[28] This iconography became standard during the reign of Ramesses II in the thirteenth century BC[27] and remained so for the rest of ancient Egyptian history, although in the Third Intermediate Period the animal forms were frequently confused.

[2] In the vignette that accompanies Spell 125 of the Book of the Dead, they appear as small figures standing on a lotus flower in front of the throne of Osiris.

[30] In the Twentieth Dynasty of the New Kingdom (1189–1077 BC), embalmers began placing wax figurines of the sons of Horus inside the body cavity.

[35] Yet in Ptolemaic and Roman times, the sons of Horus continued to appear on other burial goods, such as decorated stucco casings on wrapped mummies.

[31] A set of instructions for the embalming process, dating to the first or second century AD, calls for four officiants to take on the role of the sons of Horus as the deceased person's hand is wrapped.

[36] The last references to the sons of Horus in burial goods date to the fourth century AD, near the end of the ancient Egyptian funerary tradition.