Fig. 7

FUS depletion in neurons affects NCDN protein and mRNA levels. Lentivirus containing shRNAs towards FUS (FUS-KD1 or KD2) or non-targeted scramble (CTL) were used to infect primary rat neurons (RCN). a Confocal images of RCN (DIV16) stained with antibodies against FUS (sc-47711, red), NCDN (green) and DAPI (blue). Scale bar = 10 μm. b Bootstrapped difference of NCDN cytoplasmic intensity of medians from FUS-KD compared to CTL-KD RCN showing the kernel density plot of the bootstrapped differences (shaded grey area), the minimum and maximum resampled differences (black horizontal lines), and the 95% confidence interval (red horizontal lines) in neurons. FUS-KD1,95%CI: 0.0050- 0.1525 p = 0.0003; FUS-KD2,95% CI: 0.0057–0.1629 p = 0.001. c Western blot of NCDN, FUS and GAPDH proteins from RCN. d Quantification of NCDN protein levels from RCN relative to GAPDH. e Quantitative RT-PCR for NCDN, TDP-43 and FUS relative to U36B from primary mouse cortical neurons (DIV16). Statistical analysis was performed using a Student’s t test (a, p < 0.05; d, p < 0.001 vs CTL; ns, not significant, p > 0.05 vs CTL). Error bars represent the mean ± SEM. Each experiment was performed from n = 3–4 biological replicates per group