Figure 7.

FAK inhibition effectively abolished endothelial activation induced by CAR-T/Nalm6/PBMC co-cultured supernatant. (A) The protein level of phosphorylated FAK at tyrosine 397 as well as TF was determined by western blot. (B) The mRNA levels of endothelial activation-associated markers were determined by RT-PCR. GAPDH was taken as the housekeeping gene and data was expressed as fold changes relative to control. n = 3. (C) The protein expression of E-selectin, VCAM1, and ICAM1 was determined by flow cytometry. n = 3. (D) The concentration of IL6 and IL8 in the supernatant was determined by ELISA. (E) The concentration of Ang2 secreted by HUVEC was assessed by ELISA. n = 3. (F) Confluent HUVEC cultured in Transwell were incubated with sCAR-T supplemented with or without adalimumab for 12h. The permeability of endothelial monolayer was determined by Evans blue-BSA assay. (G) The expression levels of indicated proteins were determined by western blot. * represents p < 0.05, ** represents p < 0.01, and **** represents p < 0.0001. ns represents not significant. All data were representative of at least three independent experiments.