Virtual reality (VR) has seen tremendous advances in head-mounted displays (HMDs), optics, media quality, and other improvements that facilitate immersive experiences. With the occurrence of new technologies like Cloud VR and networked VR video services, applications such as 360° video streaming are becoming more popular within the broader consumer markets. As a result, VR content is accessible to customers with rather different levels of experiences with immersive media, i.e., never, sometimes, or often use of VR. The question, therefore, arises to which degree simulator sickness is induced to viewers depending on their experiences with VR on HMDs. In this paper, simulator sickness is evaluated for 360° videos that were shown on an HTC Vive Pro HMD to participants having different levels of experience with VR on HMDs. The modified absolute category rating with hidden reference (M-ACR-HR) method was used in a subjective experiment for video quality assessment within two subsequent sessions along with a simulator sickness questionnaire (SSQ). A statistical analysis of the SSQ scores is performed to reveal the relationship between simulator sickness and participants' experiences with VR regarding: (1) Individual symptoms, (2) Pairwise comparison of symptoms, and (3) Symptom clusters of nausea, oculomotor, disorientation, and total score. It is shown that the simulator sickness symptoms, in general, are slightly or rarely perceived across the different experience levels for the selected 360° videos. The results indicate that the reported simulator sickness increases in the second session for participants that never used VR on HMDs. Sufficiently long breaks between sessions should therefore be accounted for in the M-ACR-HR method to avoid that simulator sickness influences quality rating. © 2020 IEEE.