Fig. 3

Fkbp5 deletion attenuates LPS-induced GR activation. The HPC of WT and Fkbp5-KO mice were collected at days 1 and 7 after LPS injection, followed by mRNA and protein expression analysis. A, B qRT-PCR analysis of HPC Nr3c1 (encoding GR) and Sgk1 expression indicated that Fkbp5 deficiency reduced the LPS-induced delayed upregulation of Nr3c1 and Sgk1 expression at day 7 in the HPC. C Western blotting indicated an increase in GR on day 7 after LPS injection in the HPC of WT but not Fkbp5-KO mice. The relative band intensities were normalized to GAPDH levels. Data are indicated as the mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001 for the comparison between two groups in a two-way ANOVA followed by the Tukey post hoc test. D Immunostaining of the GR in the mouse hippocampal dorsal and ventral CA1 (dCA1 and vCA1) along with counterstaining of the cell nucleus with DAPI. GR-positive cells were predominantly located in CA1 pyramidal cell layers. E, F The nuclear localization of the GR, which indicated the activated state of the GR, was quantified by overlapping GR-DAPI signals in the total DAPI count. Note that LPS significantly increased the number of nuclear GR⁺ cells in both the dCA1 and vCA1 of WT but not Fkbp5-KO mice. ***p < 0.001 for the comparison between two groups in an unpaired Student’s t test. n = 5 mice per group. Scale bar: 50 μm