Figure 4. KXS₂₀₁₂ promotes neurogenesis in cultured neurons.

(A) KXS₂₀₁₂ (0.3 to 3 μg/mL) was applied onto cultured rat cortical neurons (DIV 5) for 96 h. The protein levels of PSD-95 (~95 kDa) and synaptotagmin (~60 kDa) were measured by Western blot assay (upper panel). α-Tubulin (~55 kDa) served as a loading control. Forskolin (Fsk, 50 nM) was used as a positive control. The amount of protein level was quantified (lower panel). (B) KXS₂₀₁₂ (3 μg/mL) and BDNF (15 ng/mL) were applied onto cultured hippocampal neurons (DIV 15) for 96 h before pEGFP-N1 transfection. The treatment significantly increased the spine density of control neurons (9–16 dendrites from 6–10 neurons per 10 μm were measured for each condition). The quantification plot was shown in lower right. (C) Representative images show that KXS₂₀₁₂ (3 μg/mL) and BDNF (15 ng/mL) increased the spine density compared to control. The counted spine density was indicated by asterisk. Scale bars = 10 μm. Values are expressed the fold of change as compared to control (x Basal) or number, Mean ± SEM, n = 4–10. *p < 0.05 and **p < 0.01 compared to the control.