Fig. 5.
Cyclooxygenase-2 (Cox-2) induction is impaired in Dgke null cells. A: quantification of Cox-2 expression, measured by real-time PCR, in bone marrow-derived macrophage (MΦ) precursors, isolated from Dgke KO mice and WT littermates and exposed to LPS or Veh as a control. Cox-2 expression is induced by stimulation with LPS in MΦ isolated from WT but not from Dgke KOs (n = 6 mice per group). AU, arbitrary units. Values are means ± SE. P values were calculated by Student's t-test. B: quantification by real-time PCR of Dgke KO and WT skin fibroblasts stimulated with IL-1β. Cox-2 expression is decreased in mutant fibroblasts and fails to be induced after IL-1β stimulation. Values are means ± SE; n = 4 mice per group. P values were calculated by Student's t-test. C: Western blot of Dgke KO and WT fibroblast lysates exposed to IL-1β or Veh as a control, probed with an antibody against Cox-2. Two replicates were loaded for each experimental condition. Cox-2 was undetectable in Dgke KO fibroblasts before and after stimulation with IL-1β, but was abundantly expressed in WT cells. For two WT samples (column 3 and 4 from left), one-half of the amount of protein was loaded to show the increase of Cox-2 expression after exposure to IL-1β. D: quantification by real-time PCR of the transcripts of Cox-2 in kidney cortexes of Dgke KO mice and WT littermates, 3 days after the injection of PAN or Veh as a control. Cox-2 expression in Dgke KO cortexes is increased in PAN-treated WT mice, but not in Dgke KOs. Values are means ± SE; n = 4 mice per group. P values were calculated by Student's t-test. E: Western blot showing the amount of Cox-2 in in kidney cortexes of Dgke KO mice and WT controls, 3 days after treatment with PAN or Veh as a control. Cox-2 is increased in cortexes of PAN-treated WT mice, but not in Dgke KOs. F: quantification by real-time PCR of Cox-2 expression in isolated glomeruli from Dgke KO mice and WT littermates, 3 days after the injection of PAN or Veh as a control. Cox-2 expression in Dgke KO glomeruli is induced in PAN-treated WT mice, but not in Dgke KOs. Values are means ± SE; n = 4 mice per group. P values were calculated by Student's t-test.