Figure 7. PGE₂ increase Bim expression during cytokine deprivation.

A. RNA was isolated from BMMC cultured in complete medium (CTRL), medium without cytokines (no cyt.) and medium without cytokines in the presence 1×10⁻⁶ M PGE₂ (no cyt. + PGE₂) for 10 h. Expression of Bim (Bcl2l11) and Puma (Bbc3), relative to GAPDH, was assessed by real time PCR using gene specific TaqMan Gene Expression Assay. B. RNA isolated as in A., Bim expression in WT, EP3−/−, and WT cells treated with ERK 1/2 inhibitor PD98059 in presence or absence of 1×10⁻⁶ M PGE₂ during cytokine deprivation. C. Expression of Bim isoform and c-Jun in WT BMMC in complete medium after stimulation with 1×10⁻⁶ M PGE₂ for indicated times. Representative of 3 independent experiments is shown. D. Expression of Bim isoforms and c-Jun various times after cytokine depletion in BMMC treated or untreated with 1×10⁻⁶ PGE₂. 1 of 6 independent experiments is shown. E. Quantitative analysis of impact of cytokine withdrawal with or without PGE₂ treatment on relative protein expression of c-Jun, Bim_S and total Bim in BMMC treated with 1×10⁻⁶ M PGE₂ during cytokines withdrawal. Data are from panels shown in C and D sections of this figure as well as from additional experiments (not shown). AU-arbitrary units, n = 6 independent experiments of 2 culture of WT and EP3−/− BMMC. Student's two-tailed t test was used to evaluate statistical differences between cytokine deprive mast cells and cytokine deprived mast cells treated with PGE₂ in A nad B. Statistical significance: * = P<0.05.