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Is it possible to run a static optimization on a model without having a .mot file? For example, I would like to apply an external load to a model to see the muscle activation to resist the load.

Please see the "Getting Started with Static Optimization" page in the documentation for a description of the required inputs (https://simtk-confluence.stanford.edu/d ... timization). Based on the information you have provided, I would suggest posing the model (using the Coordinates Panel, https://simtk-confluence.stanford.edu/d ... +and+Poses) in whatever pose you mean by "resist the load", lock all the coordinates, and run a forward dynamic simulation (the model won't move because you've locked the coordinates). You could then use the Results as the motion to track.