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. Author manuscript; available in PMC: 2021 Sep 15.
Published in final edited form as: Nat Metab. 2021 Mar 15;3(3):318–326. doi: 10.1038/s42255-021-00356-0

FIGURE 4. FKBP5 Induction Mimics FOXO4 overexpression and Fasting effects on Th Cell Activation.

FIGURE 4.

a, RT-PCR analysis of FOXO4 target genes following FOXO4 transduction in CD4+ T cells (Bar graphs - mean ± SEM from duplicate experiments in n=6 biologically independent subjects, two-sided, paired Student’s t-test). b, Representative protein blot of FKBP5 (top panel) and quantification (bottom panel) comparing transduction of control vs. FOXO4 lentivirus (OE) with (+) and without (−) CD4+ T cell activation (Bar graphs - mean ± SEM of n=6 subjects, two-sided, paired Student’s t-test). c, Cytokine release of IFNγ, IL-4 and IL-17 following FKBP5-overexpression in activated CD4+ T cells from n=9 subjects). The dot and line plots show the mean value of each subject. The values represent the average of duplicate experiments. Statistical analysis using two-sided, paired Wilcoxon test. d-e, Representative protein blot (left panel) and quantification (right panel) comparing transduction of CD4+ T cells with control vs. FKBP5 lentivirus (OE) with (+) or without (−) TCR activation (Bar graph - mean ± SEM value of subjects. d, mTOR signaling pathway phosphorylated and total protein kinase levels are shown (n=6 biologically independent experiments, two-sided, paired Student’s t-test compared to vector controls). e, STAT signaling proteins (n=8–9 biologically independent experiments, two-sided, paired Student’s t-test compared to vector controls). pSTAT1/STAT1 (n=8); pSTAT3/STAT3 (n=9); pSTAT6/STAT6 (n=9). f, Representative protein blot (top panel) and quantitative changes (bottom panel) in mouse CD4+ T cells obtained from fed- or 48-hr. fasted-mice. Protein level of FOXO4 and FKBP5 in mouse CD4+ T cells after 24-hr. TCR activation (Bar graph - mean ± SEM of n=11 mice per group, two-sided, unpaired Student’s t-test). g, The cytokine release in fed or fasted mouse CD4+ T cells. The histogram shows mean ± SEM levels from n=11 mice from duplicate assays (two-sided, unpaired Student’s t-test).

Source Data Fig.4 Unprocessed immunoblots.