Fig. 5

CRISPR/Cas9 gene editing in blood–brain barrier organoids does not disrupt barrier function. a Representative confocal images of blood–brain barrier organoids assembled with hCMEC/D3 Cas9/Scrambled gRNA control or knockout cells. Brain endothelial cells are labelled with an anti-P-gp antibody (magenta) and are homogeneously distributed at the organoid surface. Nuclei are labelled with DAPI (cyan). Scale bar, 100 μm. b Representative confocal images of blood–brain barrier organoids incubated with a non-targeting human IgG for 4 h. The upper images show an overlay of human IgG signal (yellow) and nuclei labelled with DAPI (cyan). The intensity of the lower images was scaled to visualize the background intensity in the antibody channel (grey). Scale bar, 100 μm. c Quantification of IgG intensity within blood–brain barrier organoids. Graph shows boxplots with interquartile ranges and median. Lines show the 5th and 95th percentiles. Differences between treatments were not statistically significant (p = 0.344) as evaluated by one-way ANOVA of 30 organoids per condition in n = 2 independent experiments