Figure 7.

Effect of neurospecific or ubiquitous LSD1 knockdown by shRNAs on neurite morphology in rat cortical neurons. Cultured cortical neurons were transiently transfected with pSuper GFP Neo control (scramble) (A), pSuper engineered with shRNA against exon E8a (shRNA vs 8a) (B), and pSuper engineered with shRNA against the splice junction between exons E8 and E9 (shRNA vs 8/9) (C). Morphology was analyzed for EGFP-positive neurons with DAPI (4′,6′-diamidino-2-phenylindole) counterstain. D, Cumulative neurite length in differentially LSD1 knocked down neurons is indicated as average ± SEM in micrometers. E, Secondary branches count is indicated ± SEM. F, Average neurite width by Sholl analysis calculated on inner, intermediate, and outer Sholl's circles corresponding to 20, 25, and 30 μm radii, respectively. Values shown are mean ± SEM width in micrometers. Student's t test (l Stat t I ≥ T α/2) was applied to values by comparing each condition with control scramble. *p < 0.05; **p < 0.01; ***p < 0.001. Scale bars, 20 μm.