Fig. 7.
The FICD enhances the activation of MNK1/2/eIF4E via DAP5/Fxr1. a Ingenuity pathway analysis of 145 FICD-interacting proteins. Pooled data from two biological replicates were analyzed. b Interaction map showing 20 FICD-interacting proteins in “Protein synthesis pathways” using STRING functional protein association analysis. c Frequencies of the proteins shown in (b). d The interaction of the FICD with DAP5 or Fxr1 was determined by immunoblot analysis with anti-DAP5, Fxr1, HA, or α-tubulin antibodies. Whole cell lysates of BMDMs from WT and FICDtgM mice were used for immunoprecipitation with anti-HA-tagged antibodies. Knockdown (KD) of DAP 5 (e, f, i, j) or Fxr1 (g, h, i, j) in both human CD14+ cells (e, g, i) and BMDMs (f, h, j). e–h The protein expression of NFATc1, p-eIF4E, eIF4E, DAP5, Fxr1, and α-tubulin was determined by immunoblot analysis. Osteoclastogenesis assay. KD of DAP5 or Fxr1 in hCD14+ cells (i) or in BMDMs (j) that were cultured with M-CSF and RANKL for 3 days. The left panel shows representative images of TRAP-stained cells. The right panel shows the percentages of TRAP-positive multinuclear cells (MNCs: more than three nuclei) per control from three independent experiments. CTL Control siRNAs. All data are shown as the mean ± SEM. *P < 0.05 by one-way ANOVA with a post hoc Tukey test (i, j). The data represent 2 biological replicates for mass spectrometry (a–c) and three independent experiments (d–i).