By now, the neurophysiological effect of electromagnetic field (EMF) exposure and its underlying regulating mechanisms are not well manifested. In this study, we aimed to investigate whether acute long-term evolution (LTE) EMF exposure could modulate brain functional connectivity using regional homogeneity (ReHo) method and seed-based analysis on resting-state functional magnetic resonance imaging (fMRI). We performed the LTE-EMF exposure experiment and acquired the resting-state brain activities before and after EMF exposure. Then we applied ReHo index to characterize the localized functional connectivity and seed-based method to evaluate the interregional functional connectivity. Statistical comparisons were conducted to identify the possible evidence of brain functional connectivity modulation induced by the acute LTE-EMF exposure. We found that the acute LTE-EMF exposure modulated localized intra-regional connectivity (p < 0.05, AlphaSim corrected, voxel size ! 18) and inter-regional connectivity in some brain regions (p < 0.05, AlphaSim corrected, voxel size ! 18). Our results may indicate that the approaches relying on network-level inferences could provide deeper insight into the acute effect on human functional activity induced by LTE-EMF exposure. Bioelectromagnetics. 40:42-51, 2019.