Figure 7.

Treatment with siRNA against ADAR1 compared to scrambled controls promote longer axons in rat primary cortical neurons. (A) Axons were stained with β-III tubulin (green) antibodies and actin (red) stain. (B) Bottom panel, Cortical neurons in microfluidic chambers longer axons with ADAR 1 siRNA compared to scrambled controls, Scale right panels 20 μm and microfluidic chambers in left panel 500 μm. (C) The graph is the average of 4 independent experiments in the microfluidic chambers and data points are the average with standard deviation. Statistics is t-test (*p < 0.05 and **p < 0.01) comparing Control to ADAR siRNA (blue) at a fixed distance in the microgrooves. At 800 and 900 microns, ADAR siRNA (blue) were growing significantly longer compared to controls. The graph is also displayed in Supplementary Figure 2, where we show the distribution of points for each condition. (D) In 4 independent experiments we targeted ADAR with siRNA and observed by RT-qPCR that MAP4 goes up significantly compared to scrambled controls. (E) By RT-qPCR in combination with Ribonuclease R to specifically detect circRNA, we observed up-regulation of both circRims2 (very significant) and circRTN4 (nearly significant, p-value 0.0532).