Figure 2
Figure 2. EP2 antagonism/deficiency promotes macrophage maturation. (A) EP2 antagonism promotes macrophage maturation. Freshly flushed bone marrow cells were cultured for 6 days with 30% L929 supernatant in the absence (Ctrl) or presence of EP2 antagonist AH6809 at 10μM; after 3 days, the culture was replenished with new medium containing AH6809, totaling 50% of original volume. Each day of the culture, cells were collected, stained, and analyzed for the markers indicated. Mean fluorescent intensity (MFI) is expressed as the mean ± SEM from 3 experiments, each using a different mouse (*P < .05, ***P < .001). (B) EP4 antagonism does not promote macrophage maturation. Freshly flushed bone marrow cells were cultured with the EP2 antagonist AH6809 at 10μM, or with 1μM EP4 antagonist Ono-AE3-208; control cells were cultured with DMSO. Cells were stained with F4/80-APC and CD11b-FITC and analyzed after 6 days of culture as described in panel A. Control containing DMSO was set as 100%. MFI is expressed as the mean ± SEM from 3 experiments, each using a different mouse (*P < .05). (C) Effects of aspirin (ASA) and exogenous PGE2 on macrophage maturation. Freshly flushed bone marrow cells were cultured with aspirin at 200μM, or with 1μM PGE2; control cells were cultured with DMSO. Cells were stained with F4/80-APC and CD11b-FITC and analyzed as described in panel A. Control containing DMSO was set at 100%. MFI is expressed as the mean ± SEM from 3 experiments, each using a different mouse (*P < .05, ***P < .001). (D) EP2 deficiency promotes macrophage maturation. Freshly flushed bone marrow cells isolated from EP2−/− and WT mice (wt) were cultured in 30% L929 cell supernatant as described in panel A. The dot-plot shown is representative of 3 experiments, each using cells from a different mouse. The percentage of CD11bpos/F4/80pos cells in both genotypes was compared using paired t test (*P < .05). (E) CD115 (M-CSFR) is up-regulated in EP2−/− mice during in vitro macrophage maturation. After 6 days of culture, bone marrow–derived cells were collected and analyzed for the expression of CD115 with qRT-PCR. Data are expressed as the mean ± SEM from 3 experiments, each using cells from a different mouse (*P < .05).

EP2 antagonism/deficiency promotes macrophage maturation. (A) EP2 antagonism promotes macrophage maturation. Freshly flushed bone marrow cells were cultured for 6 days with 30% L929 supernatant in the absence (Ctrl) or presence of EP2 antagonist AH6809 at 10μM; after 3 days, the culture was replenished with new medium containing AH6809, totaling 50% of original volume. Each day of the culture, cells were collected, stained, and analyzed for the markers indicated. Mean fluorescent intensity (MFI) is expressed as the mean ± SEM from 3 experiments, each using a different mouse (*P < .05, ***P < .001). (B) EP4 antagonism does not promote macrophage maturation. Freshly flushed bone marrow cells were cultured with the EP2 antagonist AH6809 at 10μM, or with 1μM EP4 antagonist Ono-AE3-208; control cells were cultured with DMSO. Cells were stained with F4/80-APC and CD11b-FITC and analyzed after 6 days of culture as described in panel A. Control containing DMSO was set as 100%. MFI is expressed as the mean ± SEM from 3 experiments, each using a different mouse (*P < .05). (C) Effects of aspirin (ASA) and exogenous PGE2 on macrophage maturation. Freshly flushed bone marrow cells were cultured with aspirin at 200μM, or with 1μM PGE2; control cells were cultured with DMSO. Cells were stained with F4/80-APC and CD11b-FITC and analyzed as described in panel A. Control containing DMSO was set at 100%. MFI is expressed as the mean ± SEM from 3 experiments, each using a different mouse (*P < .05, ***P < .001). (D) EP2 deficiency promotes macrophage maturation. Freshly flushed bone marrow cells isolated from EP2−/− and WT mice (wt) were cultured in 30% L929 cell supernatant as described in panel A. The dot-plot shown is representative of 3 experiments, each using cells from a different mouse. The percentage of CD11bpos/F4/80pos cells in both genotypes was compared using paired t test (*P < .05). (E) CD115 (M-CSFR) is up-regulated in EP2−/− mice during in vitro macrophage maturation. After 6 days of culture, bone marrow–derived cells were collected and analyzed for the expression of CD115 with qRT-PCR. Data are expressed as the mean ± SEM from 3 experiments, each using cells from a different mouse (*P < .05).

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal