Fig. 5. Preservation of blood–brain barrier integrity following AU-FUS.

a Evans Blue extravasation. Evans Blue was IV-injected. Representative IVIS spectrum images, at the FUS target location (vS1) following AU-FUS (muscimol-loaded UC-carriers with FUS, left image) vs. standard₃-FUS (right image). Regions of interests (ROIs) (1.5 ×3.5 mm, blue) were measured as radiant efficiency ipsilateral to FUS application and were compared to the contralateral vS1 (scale bar: 0.5 cm). Radiant efficiency values within ROIs for AU-FUS and standard₃-FUS sequences were quantified [n = 90 (6 rats × 15 brain sections) for AU-FUS; same rats as in Fig. 3c; n = 45 (3 rats × 15 brain sections) for standard₃-FUS]. Pairwise Mann–Whitney rank-sum test AU-FUS (ipsilateral vs. contralateral, p = 0.6577), standard₃-FUS (ipsilateral vs. contralateral, ****p < 0.0001). b MRI contrast agent extravasation. Animals were injected with Omniscan (Gd) and imaged. Representative brain images at the FUS target location (vS1) after AU-FUS (left image) compared to standard₃-FUS sequence (right image). ROIs (1.0 ×1.0 mm, orange, approximate ROI location) were measured as signal enhanced T1-weighted MR images, ipsilateral to FUS application, which were compared to the contralateral vS1 (scale bar: 1.0 cm). See Supplementary Fig. 9 for zoomed images. Baseline-subtracted contrast-enhanced T1-weighted MR image ROIs using AU-FUS and standard₃-FUS sequences were quantified [n = 9 (3 rats × 3 brain sections) for AU-FUS and standard₃-FUS]. Pairwise Mann–Whitney rank-sum test, AU-FUS (ipsilateral vs. contralateral, p = 0.9494), standard₃-FUS (ipsilateral vs. contralateral, ****p < 0.0001). c IgG immunohistochemistry. Photographic images of immunohistochemical IgG staining of brain sections from the center of the focal volume following AU-FUS (left photo) vs. standard₃-FUS (right photo) (scale bar: 0.6 cm). Normalized mean intensity values from ROI analysis of brightfield images of brain sections stained for IgG were quantified. Intensities were obtained from ROIs (1.5 × 3.5 mm) contralateral and ipsilateral to FUS treatment within the FUS focal volume [n = 27 (3 rats × 9 sections) for AU-FUS; n = 27 (3 rats × 9 sections) for standard₃-FUS] and normalized with the average contralateral mean intensity. Pairwise Mann–Whitney rank-sum test AU-FUS (ipsilateral vs. contralateral, p = 0.0970), standard₃-FUS (ipsilateral vs. contralateral, ****p < 0.0001).