Extended Data Fig. 7: Inhibiting NFAT/Calcineurin signaling reduces APOE expression and CAA pathology.

a, Chemical structures of CsA, FK506, and INCA6 showing highly dissimilar structures. b, Expression of PGK1, HPRT, and GAPDH in pericytes after two weeks with DMSO, Cyclosporine A (CsA), FK506 or INCA6. One-way ANOVA (p < 0.0001) with Bonferroni’s multiple comparison. Center values and error bars are mean expression and SD of RNA prepared from 4 independent wells for each cell line. c and d, Representative immunofluorescence imaging of APOE protein staining in pericytes after two weeks of treatment with chemicals. Scale bar, 50 μm. Experiments were repeated at least 3 times with similar results. e DEGs and associated GO terms for up-regulated and down-regulated genes in E3 and E4 CsA-treated pericyte from RNA-sequencing of RNA prepared from 3 independent wells for each condition. DEGs were determined by DSEQ2 and GO analysis was performed with Toppfun. f and g, Representative imaging and quantification depicting APOE protein expression in the APOE4KI mouse cortical slices following treatment with cyclosporine A (CsA) for one week. Unpaired, two tailed t test (p = 0.0009). Experiments were repeated with similar results using at least 3 slice preparations for each condition. Center values and error bars are mean intensity and SD from 12 independent measurements. h, Quantification of amyloid APOE4KI mouse cortical slices treated with either CsA or FK506 for one week and then exposed to 20 nM Ab for 48 hours. One-way ANOVA (p = 0.0105) with Bonferroni’s multiple comparison. Control v- CsA, p = 0.0188; FK506, p = 0.0245. Center values and error bars are mean and SD from slices prepared from 3 different mice. i, APOE mRNA expression in primary pericytes isolated from brain microvasculature of APOE4 knock-in mice treated with DMSO, Cyclosporine A, or FK506. One-way ANOVA (p = 0.0139) with Bonferroni’s multiple comparison. For DMSO v- CsA, p = 0.0221; FK506, p = 0.0367. Center values and error bars are mean and SD from pericytes prepared from 3 different mice. j, Representative image of immunostaining for APOE in hippocampal pericytes from APOE4 KI x 5xFAD mice treated with cyclosporine A or vehicle for one week. k, Representative images of vascular amyloid in the hippocampus following treatment of 6-month-old APOE4KI x 5XFAD female mice with either vehicle or CsA. Amyloid was detected and quantified with two independent anti-amyloid antibodies (6e10 and 12F4). These experiments were repeated 2 times with similar results.