Ultrasonography was used to measure pennation angle and electromyography (EMG) to record muscle activity of the human tibialis anterior (TA), lateral gastrocnemius (LG), medial gastrocnemius (MG), and soleus (SOL) muscles during graded isometric ankle plantar and dorsiflexion contractions done on a Biodex dynamometer. Data from eight male and eight female subjects were collected in increments of approximately 25% of maximum voluntary contraction (MVC) ranging from rest to MVC. A significant positive linear relationship (p < 0.05) between normalized EMG and pennation angle for all muscles was observed when subject specific pennation angles at rest and MVC were included in the analysis. These were included to account for gender differences and inter-subject variability in pennation angle. The coefficient of determination, R 2 , ranged between 0.76 for the TA to 0.87 for the SOL. The EMG-pennation angle relationships have ramifications for use in EMGdriven models of muscle force. The regression equations can be used to characterize fiber pennation angle more accurately and to determine how it changes with contraction intensity, thus providing improved estimates of muscle force when using musculoskeletal models.