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Microtubule-kinesin active fluids consume ATP to generate internal active stresses, driving spontaneous and complex flows. While numerous studies have explored the fluid's autonomous behavior, its response to external mechanical forces remains less understood. This study explores how moving boundaries affect the flow dynamics of this active fluid when confined in a thin cuboidal cavity. Our experiments demonstrate a transition from chaotic, disordered vortices to a single, coherent system-wide vortex as boundary speed increases, resembling the behavior of passive fluids like water. Furthermore, our confocal microscopy revealed that boundary motion altered the microtubule network structure near the moving boundary. In the absence of motion, the network exhibited a disordered, isotropic configuration. However, as the boundary moved, microtubule bundles aligned with the shear flow, resulting in a thicker, tilted nematic layer extending over a greater distance from the moving boundary. These findings highlight the competing influences of external shear stress and internal active stress on both flow kinematics and microtubule network structure. This work provides insight into the mechanical properties of active fluids, with potential applications in areas such as adaptive biomaterials that respond to mechanical stimuli in biological environments. *We acknowledge support from the National Science Foundation (NSF-CBET-2045621). This research is performed with computational resources supported by the Academic & Research Computing Group at Worcester Polytechnic Institute. We acknowledge the Brandeis Materials Research Science and Engineering Center (NSF-MRSEC-DMR-2011846) for use of the Biological Materials Facility. 

Active fluids with spatiotemporally varying activity have potential applications to micromixing; however previously existing active fluids models are not prepared to account for spatiotemporally-varying active stresses. Our experimental work used UV-activated caged ATP to activate controlled regions of microtubule-kinesin active fluid inducing a propagating active-passive interface. Here, we recapitulate our experimental results with two models. The first model redistributes an initial ATP distribution by Fick's law and translates the ATP distribution into a velocity profile by Michaelis-Menton kinetics. This model reproduces our experimental measurements for the low-Péclet number limit within 10% error without fitting parameters. However, as the model is diffusion based, it fails to capture the convective based superdiffusive-like behaviour at high Péclet numbers. Our second model introduces a spatiotemporally varying ATP field to an existing nematohydrodynamic active fluid model and then couples the active stresses to local ATP concentrations. This model is successful in qualitatively capturing the superdiffusive-like progression of the active-inactive interface for high Peclet number (convective transport) experimental cases. Our results show that new model frameworks are necessary for capturing the behaviour of active fluid with spatiotemporally varying activity. *T.E.B., E.H.T., J.H.D., and K.-T.W. acknowledge support from the National Science Foundation (NSF-CBET-2045621). C.-C. C. was supported through the National Science and Technology Council (NSTC), Taiwan (111-2221-E-006-102-MY3). M.M.N. was supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences (DE-SC0022280). 

Abstract Active fluids have applications in micromixing, but little is known about the mixing kinematics of systems with spatiotemporally-varying activity. To investigate, UV-activated caged ATP is used to activate controlled regions of microtubule-kinesin active fluid and the mixing process is observed with fluorescent tracers and molecular dyes. At low Péclet numbers (diffusive transport), the active-inactive interface progresses toward the inactive area in a diffusion-like manner that is described by a simple model combining diffusion with Michaelis-Menten kinetics. At high Péclet numbers (convective transport), the active-inactive interface progresses in a superdiffusion-like manner that is qualitatively captured by an active-fluid hydrodynamic model coupled to ATP transport. Results show that active fluid mixing involves complex coupling between distribution of active stress and active transport of ATP and reduces mixing time for suspended components with decreased impact of initial component distribution. This work will inform application of active fluids to promote micromixing in microfluidic devices.