The driving conduct is a key factor with respect to both operational and energetic performance of railway vehicles. In this respect, a Hardware-in-the-Loop (HiL) test bench has been designed and set-up. It is based on a main virtual simulation environment for railway operation, that includes two vehicles running on the same track; in addition, an external physical board is present, that contains the on-board unit of one vehicle, and allows addressing communication issues; to run the simulations in real-time and provide analog input and output ports, a Speedgoat® target machine is included, and it is connected to a physical driving interface (made of two handles), that emulate the dashboard of a real train. This HiL test bench configuration allows to evaluate different driving styles considering various performance indicators, such as punctuality and energy efficiency. The virtual simulation environment may include different signalling systems, based on Fixed Block (FB) and Moving Block (MB) approaches, so that it allows investigating the combined effect of different driving strategies and signalling logics. Two features characterize the driving conduct, namely defensiveness and reactiveness: the former determines how smoothly the driver modulates traction/braking effort respectively during acceleration/deceleration phases; the latter is related to how fast the driver responds to inputs received from the on-board equipment, such as a warning signals. Different combinations of these two characteristics give rise to a wide range of driving strategies, with the aim of finding relevant trends between driving conducts and performance levels, in order to provide repeatable instructions for optimal manual driving.