The influence of molecular structure on the mechanical properties of self-assembled 1,3,5-benzenetrisamide nanofibers is investigated. Three compounds with different amide connectivity and different alkyl substituents are compared. All the trisamides form well-defined fibers and exhibit significant differences in diameters of up to one order of magnitude. Using nanomechanical bending experiments, the rigidity of the nanofibers shows a difference of up to three orders of magnitude. Calculation of Young's modulus reveals that these differences are a size effect and that the moduli of all systems are similar and in the lower GPa range. This demonstrates that variation of the molecular structure allows changing of the fibers' morphology, whereas it has a minor influence on their modulus. Consequently, the stiffness of the self-assembled nanofibers can be tuned over a wide range-a crucial property for applications as versatile nano- and micromechanical components.