Figure 1.

ASOs taken up via gymnosis are active in neurons, microglia, and astrocytes following i.c.v. injection

(A) C57BL/6N mice were i.c.v. injected with saline or 100 μg TA1, and analyzed over 14 days. Illustration created using BioRender.com. (B) Acute toxicity 1 h post injection. (C) Percent body weight change normalized to day 0, showing no acute toxicity over the course of the experiment. Error bars cannot be visualized as plotted. (D and E) Representative IF images of brain sections stained for TA1. (D) Coronal brain section stained 2 days post injection of 100 μg TA1. Scale bars, 2 mm and 500 μm (inset). (E) TA1 colocalizes with NeuN (left) as well as Iba1 (right) 14 days post injection of 100 μg TA1. Scale bars, 100 μm. (F–J) Neurons, microglia, and astrocytes were FACS sorted and analyzed for TMEM106b mRNA expression levels 14 days post saline (n = 4) or TA1 (100 μg, n = 4) i.c.v. injection. (F) Gating strategy for cell-specific FACS sorting. (G–J) Relative TMEM106b mRNA expression levels in unsorted brain (G), and sorted neurons (H), microglia (I), or astrocytes (J) as assessed by qRT-PCR. Data were normalized to actin- and saline-treated samples. For all panels, mean ± SEM are shown. Unpaired two-tailed Student’s t test, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.