Figure 2.

Immune response following in vivo editing with viral and non-viral Cas9 delivery strategies

(A) Representative immunostaining of Iba1 (microglia, green) with tdTomato and DAPI using confocal microscopy. Scale bar, 50 $\mu$m. (B) Quantification of Iba1⁺ staining intensity and percent area (n = 4–6 technical replicates, one-way ANOVA, ∗p < 0.05). (C) Quantification of CD45⁺ and CD3⁺ cells per image (n = 3–6 replicates, one-way ANOVA, ns). (D) Representative images of CD45, CD3, and Iba1 showing co-expression of CD45 (green) with both Iba1 (microglia, red) and CD3 (T cells, red) cells and differential cell morphology. Merged images include DAPI (gray) and tdTomato (magenta). Scale bars, 50 $\mu$m. (E) Quantification of IgG antibodies against Cas9 or AAV capsid proteins measured 28 and 90 days after bilateral intrastriatal injections by ELISA (n = 3–5 biological replicates). (F) Heatmap summarizing qRT-PCR results of gene expression from homogenized brain tissue near the injection site (striatum and cortex) at two time points. Ppih was used as a housekeeping control for delta-delta Ct analysis and compared with the sham group using the QIAGEN analysis portal (n = 4, ∗p < 0.05).