Janusene is a symmetrical molecule that contains four benzene rings, with two of them forced to be in a vertical quasi-parallel face-to-face alignment. The unique physical nature of the transannular interactions and the electronic features of the region between the enforced parallel rings was tested with the complexation of Ag⁺ ion as a probe to evaluate the interplay between π-stacking and cation–π non-bonded interactions. The janusene framework and the [janusene–Ag]⁺ host–guest (H–G) system were analyzed through the introduction of substituent groups with different chemical natures and in different parts of the host framework. The janusenes were used to tune both π-stacking and cation–π interactions. Three modes of substitution (facial, lateral, and facial plus lateral) were explored to gain insight into the effects of such scaffold modifications on the dual non-bonded interactions. Our findings suggest that the η²:η² silver coordination is the most stable interaction mode between the silver ion and the janusene parallel rings. The cation–π interaction in the host structure is stabilized by electron donating groups and destabilized by electron withdrawing groups. The stabilization effect is highlighted with substitutions on the facial and facial plus lateral modes, with the latter being due to additive cooperation between the substituent groups. The bonding analysis indicates that [janusene–Ag]⁺ complexes containing electron withdrawing groups in the facial and facial plus lateral substitution schemes are more stabilized by orbital interactions. Complexes with electron donating groups and the complexes with substituent groups in the lateral position are mainly stabilized by electrostatic interactions, although in all cases orbital and dispersive interactions are also essential to describe the bonding situation. We envisage that these results will guide the development of new systems with increased cation–π interaction capability.