The reliability of nanocomposite conductive inks under cyclic loading is the key to designing robust flexible electronics. Although resistance increases with cycling and models exist, the exact degradation mechanism is not well understood and is critical for developing inks. This study links cracking behavior to changes in electrical resistance by performing in situ cyclic stretch experiments in scanning electron microscopy (SEM) with synchronized resistance measurements. Two screen-printed conductive inks, PE874 and 5025, on thermoplastic polyurethane (TPU) and polyimide (PI) substrates, respectively, were tested using the in situ technique. The obtained SEM images were analyzed with digital image correlation (DIC) to map the strain across cycles. The strain maps show that fatigue damage mainly occurred within the cracks formed during the initial monotonic stretch. There was no delamination at the ink−substrate interface or crack extension along the surface with cycling. Instead, fatigue damage resulted from a combination of crack widening and local shearing within the existing cracks. Crack depth varied based on the ink and substrate properties. The cracks in the 5025 ink on the PI substrate were only partially through the ink thickness, while fully through-thickness cracks were more prevalent in the PE874 ink on the TPU substrate. The 5025 ink showed a faster resistance increase with cycling than the PE874 ink because fatigue damage affected more bridging ink material for partial through-thickness cracks. Higher strain amplitudes caused greater crack widening and shearing and therefore faster resistance increase per cycle.