Abstract. The neurovisceral integration model proposes that different forms of self-regulation, including the emotional suppression, are characterized by the activation of neural network whose workings are also reflected in respiratory sinus arrhythmia (RSA). However, most of the previous studies failed to observe theoretically expected increases in RSA during emotional suppression. Even when such effects were observed, it was not clear whether they resulted from specific task demands, a decrease in muscle activity, or they were the consequence of more specific self-control processes. We investigated the relation between habitual or trait-like suppression, spontaneous, and instructed suppression with changes in RSA during negative emotion experience. A modest positive correlation between spontaneous situational and habitual suppression was observed across two experimental tasks. Furthermore, the results showed greater RSA increase among participants who experienced higher negative affect (NA) increase and reported higher spontaneous suppression than among those with higher NA increase and lower spontaneous suppression. Importantly, this effect was independent from the habitual suppression and observable facial expressions. The results of the additional task based on experimental manipulation, rather than spontaneous use of situational suppression, indicated a similar relation between suppression and RSA. Our results consistently demonstrate that emotional suppression, especially its self-regulation component, is followed by the increase in parasympathetic activity.