Krüppel-like factor 9 (Klf9) regulates fkbp5. (A) Sinusoid modeling of fkbp5 time-course data obtained by quantitative reverse transcription and polymerase chain reaction (qRT-PCR) of RNA extracted from wildtype (WT) and klf9–/– larvae. Each data point represents the average of three biological replicates of pooled larvae. P-value calculated by paired t-test (data paired at each timepoint). Error bars represent standard error of the mean. Model fits tested by ANOVA (P < 0.0001, see Supplementary Figure 4). (B) Distribution of Midline Estimated Statistics Of Rhythmicity (MESORs) of datasets generated by random sampling as described in Figure 2 and Methods. MESORs of measured WT and klf9–/– data sit at either extreme. (C) Schematic of putative Klf9 target motifs in the fkbp5 promoter region. Black arrows indicate sites of primers used in panels (D,E). (D) Recovery of fkbp5 promoter DNA from WT (-) and Klf9-AM tagged (+) larvae by ChIP with anti-AM tag antibody. (E) Fold enrichment (FE) of fkbp5 promoter DNA recovered in klf9–/– mutants with ChIP using antibody to acetyled-H3K14. (F) Proposed gene regulatory circuit containing the glucocorticoid receptor (GR), fkbp5, and klf9. Activation is denoted by green arrows, repression by red lines.
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