Fig. 1: Effects of (R)-ketamine and (S)-ketamine on the BDNF activation.

A, B The luciferase assay for BDNF exon IV promoter. A BDNF exon IV promoter activity in the HEK293T cells treated with (R)-ketamine (0.1, 1.0, and 10 μM) or (S)-ketamine (0.1, 1.0, and 10 μM). The data are the mean ± SEM (n = 6). **P < 0.01; ***P < 0.001 compared to vehicle group. ^$P < 0.05; ^$$P < 0.01; ^$$$P < 0.001 (one-way ANOVA). B BDNF exon IV promoter activity in the HEK293T cells treated with (R)-ketamine (10 μM) [or (S)-ketamine (10 μM)] with or without mutation (Mut) plasmids. The data are the mean ± SEM (n = 9 or 10). **P < 0.01; ***P < 0.001 (one-way ANOVA). C BDNF exon IV promoter activity in the HEK293T cells treated with (R)-ketamine (10 μM) with or without siRNA-CREB plasmids. The data are the mean ± SEM (n = 4). **P < 0.01; ***P < 0.001 (one-way ANOVA). D BDNF exon IV promoter activity in the HEK293T cells treated with (R)-ketamine (10 μM) with or without CREB-HDO or BDNF exon IV-HDO. The data are the mean ± SEM (n = 10). **P < 0.01; ***P < 0.001 (one-way ANOVA). E ChIP-PCR assay for BDNF IV promoter. The p-CREB protein–DNA crosslinking samples were obtained from the primary microglia treated with vehicle, (R)-ketamine (10 μM), or (S)-ketamine (10 μM) via co-immunoprecipitating with anti-p-CREB. PCR was carried out by using BDNF exon IV promoter primer. The data are the mean ± SEM (n = 4). *P < 0.05; ***P < 0.001 (one-way ANOVA). F The immunofluorescence for p-CREB and MeCP2 in the LPS (1 µg/mL)-treated primary microglia with (R)-ketamine (10 µM) or (S)-ketamine (10 µM). Scale bar = 50 μm. G Expression of NRBP1, BDNF, MeCP2, and the ratio of p-CREB/CREB in the LPS-treated primary microglia with (R)-ketamine (10 µM) or (S)-ketamine (10 µM). The data are the mean ± SEM (n = 5). *P < 0.05; **P < 0.01; ***P < 0.001 (one-way ANOVA).