Fig. 5. Effect of the intra-arterial administration of miR-96-5p inhibitor with MBs and US on the levels of GSH, EAAC1, GTRAP3-18, and NOVA1.

a A schematic of the intra-arterial (i.a.) injection is shown. The common carotid artery (CCA), external carotid artery (ECA), and internal carotid artery (ICA) are exposed, and a small incision is made on the CCA. The catheter is inserted and advanced towards the ICA; once the catheter is stably positioned, a mixture of miR-96-5p inhibitor and MBs is injected with a programmed microinjector. Simultaneously with the injection, ultrasound (US) is administered from a probe placed on the shaved heads of the mice. b Confocal images show the effect of administration of a miR-96-5p inhibitor or a negative control (NC) inhibitor on the intensity of CMFDA as an intracellular GSH marker (green). NeuN is stained as a neuronal nuclear marker (red) and co-localized with the CMFDA signal. Scale bar, 50 μm. c Confocal images show the effect of administration of a miR-96-5p inhibitor or an NC inhibitor on the expressions of GTRAP3-18 (green) and NOVA1 (red). The nuclei were stained with DAPI (blue). Scale bar, 50 μm. d Confocal images show the effect of administration of a miR-96-5p inhibitor or an NC inhibitor on the EAAC1 expression (green). The nuclei were stained with DAPI (blue). Scale bar, 50 μm. e The level of CMFDA intensity after the administration of NC or miR-96-5p inhibitor are shown. Data are mean values ± SD obtained from four independent experiments and individual data points are plotted. Data were analyzed by Student’s t-test, two-sided. *p < 0.05 relative to NC inhibitor. f–h The expression of EAAC1 (f), GTRAP3-18 (g), and NOVA1 (h) after the administration of NC or miR-96-5p inhibitor are shown. Data are mean values ± SD obtained from independent samples (n = 3 for NC inhibitor, n = 4 for miR-96-5p inhibitor) and individual data points are plotted. Data were analyzed by Student’s t-test, two-sided. *p < 0.05 relative to NC inhibitor.