Fig. 6

The impact of RN7SL1 knockdown on neuronal apoptosis and morphology through microglia. (a) After stimulating primary neurons with primary microglial conditioned medium (CM) for 24 h, representative images depict apoptotic neurons identified by TUNEL assay (red) alongside NeuN staining (green; neuronal marker). Statistical results represent the ratio of TUNEL⁺ cells to NeuN⁺ cells (n = 6/group). (b) After stimulating primary neurons with primary microglial CM for 24 h, representative images were captured of the neurons immunostained with MAP2 (red) and 4′,6-diamidino-2-phenylindole (DAPI; blue). Three-dimensional reconstruction of the neurons was conducted using Imaris. Statistical analyses included dendrite branch level (n = 8/group) and neuron quantification through Sholl analysis (n = 8/group). (c) After stimulating primary neurons with primary microglial CM for 24 h, representative confocal microscopy images show the dendritic spines of the neurons immunostained with phalloidin (green) for spine visualization and MAP2 (red) for neuronal structure. Statistical analysis was conducted on dendritic spine density and length (n = 6/group). Values represent mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, as determined by one-way ANOVA.