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. 2022 Aug 2;11(15):2384. doi: 10.3390/cells11152384

Figure 6.

Figure 6

Blockade of 5-HT2A with ketanserin reversed TrkB autophosphorylation in vitro and in vivo. (A) Neuroblastoma N1E-115 cells expressing HA-tagged 5-HT2A receptor, GFP-tagged TrkB, or co-expressing both receptors were treated with the 5-HT2A receptor antagonist ketanserin (1 μM) for 10 min, followed by treatment with either 7,8-DHF (500 nM) or vehicle for 30 min. Cell lysates were then subjected to SDS-PAGE and Western blot (WB) analysis with antibodies against phosphorylated or total TrkB. β-tubulin was used as a loading control. Representative WBs are shown. Bars show means ± SEM (n ≤ 8). * p < 0.05; *** p < 0.001 (two-way ANOVA). (B) Mice were intraperitoneally injected with ketanserin (1 mg/kg) or 25CN-NBOH (1 mg/kg), resulting in increased TrkB phosphorylation in the hippocampus and frontal cortex. At 10 min (for 25CN-NBOH) or 30 min (for ketanserin) after injection, the animals were euthanized, and brain lysates were prepared from the frontal cortex, hippocampus, and striatum. This was followed by SDS-PAGE and WB to detect phosphorylated and total TrkB. GAPDH was used as a loading control. Representative WBs are shown. Bars represent means ± SEM (5 ≤ n ≤ 8) * p < 0.05; ** p < 0.01 (two-way ANOVA).