Experimental design

Experimental animal studies were performed in accordance with the Animal Research: Reporting of in vivo Experiments (ARRIVE) guidelines with government approval from the State Agency for Nature, the Environment and Consumer Protection North Rhine-Westphalia. The procedure used for the laboratory animals was approved by the Institutional Animal Care and Use Committee (IACUC) of Shanghai Jiao Tong University, China. Animals were housed in cages under standard laboratory conditions. The experimental schedule is shown in Figure 1. Adult male ICR mice underwent 60 minutes of transient MCAO. Occlusion was confirmed by the reduction of cerebral blood flow, as determined by a laser Doppler flowmetry. One week after ischemia, 0.5 MHz low frequency tFUS was applied to the ischemic hemisphere of mice for 7 consecutive days. Brain atrophy volume was calculated via cresyl violet staining. Neurobehavioral outcomes were assessed by neurological severity score (NSS), elevated body swing test (EBST) and corner test. IL-10 and IL-10R were examined to explore the underlying mechanism.

Flow diagram of the experimental design Animals underwent middle cerebral artery occlusion (MCAO). tFUS was applied for 7 days from day 7 of MCAO. The behavioral tests were performed following MCAO for 14 days. Brain atrophy volume and potential mechanisms were examined.