Not DOS, but two similar era cases.

A FTSE 100 company I used to work for, still maintains a SCO UNIX machine that was first installed in 1993. It hasn't had any upgrades since, has specialist hardware in it, and still has an IP stack that is classful (ie. no subnet masks, 10.x.x.x is always a /8). It runs in a 100% airgapped network, and it isn't essential to keeping the core companies operations, but it is a ticking time bomb nobody seems to want to address that will cause a lot of distruption when it finally dies.

Also I was recently (very lightly) involved in a consultancy project to remove an OS/2 based point of sale system for a small local retailer with 6 stores.