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Automation misuse and acceptance, influenced by trust, environmental conditions, and confidence, have hindered drivers from fully benefiting from partially automated vehicles. This study investigates how driver trust changes with AV reliance, differences in mental and physiological states, and continuous measures’ effectiveness. The takeover drivers reported lower trust than the non-takeover drivers in all scenarios. Nontakeover drivers’ elevated DLPFC activation aligns with trust networks and emotion regulation. The groups also differed in neural activation preand during scenarios with the takeover group showed more PFC, V2V3, and IFC engagement pre-scenario. Gaze revealed the takeover group fixated more on the AV button or dashboard, indicating readiness to take over, while non-takeover drivers focused on the rearview mirror, reflecting situational awareness. HRV responses showed higher physiological arousal in the takeover group pre-scenario. In summary, our multimodal approach reveals takeover behavior is associated with lower trust, cognitive unloading, increased stress, and anticipatory visual attention.