Fig. 6

Loss of NCDN affects FUS localization and solubility. Lentivirus containing shRNAs targeting NCDN (NCDN-KD1 or -KD2) or non-targeted scramble (CTL-KD) were used to deplete Neuro-2a cells of NCDN. a CTL-KD and NCDN-KD cells were lysed in PLB followed by fractionation to generate soluble (S1) and insoluble fractions (P1). TCL: total cell lysates. PLB: polyribosome lysis buffer. RIPA: radioimmunoprecipitation assay buffer. b Western blot of NCDN, FUS and GAPDH proteins and corresponding ponceau red staining of membranes. 5% of each fraction was loaded on the gel. The arrow indicates the band corresponding to NCDN. c Quantification of FUS protein levels in fractions. Statistical analysis was performed using a Student’s t test (a, p < 0.05; b, p < 0.01; 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