Data Sheet 1_Focused ultrasound increases Lucifer Yellow transport across hCMEC/D3 endothelial monolayers.pdf

Lucifer Yellow accumulation was quantified immediately before exposure, at the end of exposure, and 9 min later in eight independent paired experiments using sampling- and volume-corrected receiver amounts.</p>Results<p>During the 60-s exposure interval, the mean control-adjusted increase was 0.283 nmol (95% confidence interval, 0.185–0.380 nmol; Holm-adjusted P=0.000475; Cohen’s dz=2.43), and all eight experiments showed the same effect direction.

During the subsequent 9-min interval, the additional control-adjusted accumulation was 0.035 nmol (95% confidence interval, −0.100–0.171 nmol; Holm-adjusted P=0.555). In a separate preliminary cell-free session, three within-session FUS/control pairs did not reproduce the positive exposure-interval response seen in the cellular series; these observations were descriptive only. In five separate paired experiments, acute Calcein-AM- and ethidium homodimer-1-derived image metrics showed mixed FUS-minus-control differences without a reproducible FUS-associated direction.</p>Discussion<p>These findings demonstrate an exposure-associated increase in Lucifer Yellow transport across the hCMEC/D3 monolayer–insert system that arose predominantly during insonification, without statistically detectable continued differential accumulation during the observed post-exposure interval.

Together, these results provide a controlled in vitro framework for temporally resolving FUS-associated transport in this monolayer–insert system.</p>