Figure 1: FKBP51 directs autophagic pathways.
(B) Change of total Beclin1, pAktS473, and pAktT308 upon expression of FKBP51. Representative Western blots on the right. FKBP51 was detected by an antibody directed against its FLAG tag.
(C) FKBP51 interacts preferentially with dephosphorylated Akt and promotes Akt dephosphorylation. Graphs display quantification of pBeclin1 (S234 and S295) and Beclin1-Akt/pAktS473 interaction after Beclin1 immunoprecipitation from HEK cells transfected with FKBP51/52 (details in Figure S2).
(D) Comparison of the endogenous protein levels of effects of Beclin1, LC3B-II/I, Atg12, and pAktS473 levels in wild-type MEFs, 51KO MEFs, and 51KO MEFs transfected with FKBP51 or vector control. Graphs show the relative expression, with the levels of wild-type cells transfected with vector control set to 1 (dashed line). Representative Western blots on the right.
(E) Quantification of Beclin1-Akt interaction after Beclin1 immunoprecipitation in brain extracts from wild-type and 51KO mice. Graph represents the results from five independent experiments. Representative Western blots on the right.
(F) FKBP51 enhances autophagic flux. Cortical rat astrocytes were transfected with FKBP51 or vector and treated with bafilomycin A1 (BafA1) as indicated, and the levels of LCB3-II/I were determined.
(G) Cellular effects of FKBP51 on autophagic markers require Akt1 and/or Akt2. The protein levels of Beclin1 and LC3B-II/I were evaluated in wild-type MEFs, Akt1/2KO MEFs, and Akt1/2KO MEFs transfected with Akt1- and Akt2-expressing plasmids. Graphs display the relative expression of three different experiments; expression in wild-type vector-transfected MEFs was set to 1 (dashed line). Representative Western blots on the right. *p<0.05; **p<0.01; ***p<0.001. See Table S1 for statistical details. ect., ectopic; genot., genotype; KO, 51KO; WT, wild-type.
Cited from: http://dx.doi.org/10.1371/journal.pmed.1001755.g002
Figure 2: Model of FKBP51’s impact on Beclin1 and autophagy pathways.
FKBP51 interacts with PHLPP, Akt, and Beclin1. Since PHLPP dephosphorylates Akt (S473 in Akt1), inactive Akt is recruited to Beclin1. This results in lower phosphorylation of Beclin1 and, thus, the induction of autophagic pathways [19]. Autophagic pathways, and thereby also FKBP51, are linked to cell homeostasis and to synaptic (syn.) function [66] as a physiological correlate of behavior [71]. Antidepressants act on the same pathways in an FKBP51-dependent manner and change FKBP51 protein interactions; this could form the basis for the FKBP51 dependency of antidepressant effects in cells, animals, and humans. Cited from: http://dx.doi.org/10.1371/journal.pmed.1001755.g010
Figure 3: Schematic of study design. (doi:10.1371/journal.pmed.1001102.g001)
Figure 4: Schematic overview of experimental workflow. PBMCs were either gamma-irradiated with 60 Gy or not irradiated. After culturing for the indicated times, cells were centrifuged to separate the cell pellet and CM supernatant. The pellet was used to extract cellular RNA, which was used for microarray analysis. The CM supernatant was processed to separate and isolate different molecular components. The steps highlighted in blue indicate samples used for in vitro assays. The different methods and bioinformatics tools used for sample analyses are indicated at the appropriate links.
Cited from: Scientific Reports 5, Article number: 16662 (2015). doi:10.1038/srep16662
Figure 5: CP13 immunocytochemistry.
….. Images as shown were analyzed with ImageJ for percent area stained. Each data point represents the mean value obtained from at least two sections per mouse. The area stained is significantly lower from CP13 treated mice (p < 0.009) and significantly higher in PG5 treated mice (p < 0.019).
doi:10.1371/journal.pone.0135774.g006
