Fig. 3

TRPV4 regulates barrier resistance and Cldn5 expression in human brain ECs. a TRPV4 measured in knock down of TRPV4 (shTRPV4) compared to non-targeting shRNA control human brain ECs (NTC), N = 5. b Protein levels of TRPV4 in shTRPV4 cells compared to NTC cells. c TEER measurement of shTRPV4 and NTC brain ECs and quantification of max. resistance normalized to cell attachment (t = 0), N = 5. d mRNA expression of CLDN5, VE-Cad and ZO-1 in shTRPV4 and NTC cells, N = 8. e Representative images of Cldn5 immunoreactivity in shTRPV4 cells and NTC cells, orange arrowheads indicate cell–cell junctions; scale bar: 25 µm. f Protein levels of TRPV4 in TRPV4 OE compared to EV cells. g Calcium baseline and TRPV4 agonist-mediated calcium response in TRPV4 OE compared to EV cells (GSK1016790A, 100 nM). Quantification of TRPV4 agonist-mediated calcium response, N = 3. h TEER measurement of TRPV4 OE and EV brain ECs and quantification of max. resistance normalized to t = 0, N = 3. i Representative images of Cldn5, VE-Cad, and F-actin immunoreactivity in TRPV4 OE and EV cells, orange arrowheads indicate cell–cell junctions; scale bar: 10 µm. Data is shown as mean ± SEM and an average of technical replicates in each biological replicate. Statistics were performed by paired Student’s t-test indicated by connecting lines (*p < 0.05)