Fig. 6

LIPUS enhances the proportion of neuroprotective astrocytes with the help of neuron-secreted Irisin. Primary hippocampal astrocytes were harvested from C57BL/6J mouse pups within 24 h after birth. A Primary hippocampal astrocytes at DIV 12 were treated with LIPUS and then harvested 2 days later. Cell viability was assessed using CCK-8 Kit. B, C Primary hippocampal astrocytes at DIV 12 were injured by OGD for 6 h and then harvested 2 days later. Fndc5 expression and S100A10 (neuroprotective phenotype marker) level were assessed by qPCR between the astrocytes with OGD injure and the astrocytes with OGD injure as well as LIPUS treatment. D– F Primary hippocampal astrocytes at DIV 12 were replaced medium with the NCM, simultaneously processed with 6 h OGD as well as 2 day LIPUS, and then then harvested 2days later. S100A10 and GFAP (reactive astrocyte marker) were used to determine neuroprotective astrocyte (S100A10 and GFAP co-labeled cell), the representative images of astrocytes in immunohistochemistry (D). The proportion of neuroprotective astrocytes were detected using the ImageJ software (E). The level of TGF-β in the supernatants of astrocytes were determined using ELISA (F). (n = 6 per group, 6 independent experiments from 6 different astrocytes preps. All data are expressed as mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001, ns means no statistical significance)