PPARγ exerts significant anti-inflammatory signaling properties in monocytes and macrophages, which are affected by its intracellular localization. Based on our previous report, which showed that cytosolic localization of PPARγ attenuates PKCα signaling in macrophages, we elucidated the molecular mechanisms provoking cytosolic PPARγ localization. Using the DsRed-tagged PPARγ deletion constructs PPARγ1 Δ1-31 and PPARγ1 Δ407-475, we observed an exclusive nuclear PPARγ1 Δ1-31 localization in transfected HEK293 cells, whereas PPARγ1 Δ407-475 did not alter its cytosolic or nuclear localization. The casein kinase II (CK-II) inhibitor 5,6-dichloro-1-β-D-ribofuranosyl benzimidazole (DRB) prevented cytosolic PPARγ localization. Mutation of two possible CK-II phosphorylation sites at serine 16 and serine 21 of PPARγ into alanine (PPARγ S16A/S21A) inhibited cytosolic PPARγ localization. Moreover, a PPARγ S16E/S21E mutant that mimicks constitutive phosphorylation of residues 16 and 21, predominantly resides in the cytosol. The CRM1 inhibitor leptomycin B abolished cytosolic PPARγ localization, suggesting that this is a CRM1-dependent export process. CRM1-mediated PPARγ export requires Ran and phosphorylated RanBP3. Finally, co-immunoprecipitation studies demonstrated that DRB blocks PPARγ binding to CRM1, whereas PD98059 inhibits RanBP3 binding to CRM1 and concomitant shuttling from nucleus to cytosol, but does not alter PPARγ binding to CRM1. We conclude that CK-II-dependent PPARγ phosphorylation at Ser16 and Ser21 is necessary for CRM1/Ran/RanBP3-mediated nucleocytoplasmic translocation of PPARγ.