Figure 3
Figure 3. GCs induce phosphorylation of ERK1/2, but not JNK1/2 and p38. Monocytes were stimulated with 100nM DEX (GC) or medium as a control (Co) for 12, 24, and 48 hours. Afterward, cells were lysed, and Western blot analysis was performed with antibodies recognizing phosphorylated and unphosphorylated forms of ERK1/2, JNK1/2, and p38 MAP kinases. The figure shows immunoblots from one of 3 representative experiments (A). Monocytes were stimulated with DEX (GC) and medium as control (Co) or resuspended in conditioned media from control (SN Co) or GC-treated monocytes (SN GC) and cultured for 24 hours. The figure shows immunoblots for analysis of ERK phosphorylation from 1 of 3 representative experiments (B). Monocytes were pretreated for 30′ in the presence of 3μM RU486 before 16 hours stimulation with DEX (GC) or medium as a control (Co). Subsequently, cells were stimulated to undergo apoptosis by STS (■) or left untreated (□). After 6 hours, the proportion of annexin V–positive cells was measured (C). Data show mean ± SEM (n = 3). In parallel, cells were lysed, and immunoblotting was performed using antiphosphorylated and total ERK1/2 antibodies (D). The figure shows immunoblots from 1 of 3 representative experiments.

GCs induce phosphorylation of ERK1/2, but not JNK1/2 and p38. Monocytes were stimulated with 100nM DEX (GC) or medium as a control (Co) for 12, 24, and 48 hours. Afterward, cells were lysed, and Western blot analysis was performed with antibodies recognizing phosphorylated and unphosphorylated forms of ERK1/2, JNK1/2, and p38 MAP kinases. The figure shows immunoblots from one of 3 representative experiments (A). Monocytes were stimulated with DEX (GC) and medium as control (Co) or resuspended in conditioned media from control (SN Co) or GC-treated monocytes (SN GC) and cultured for 24 hours. The figure shows immunoblots for analysis of ERK phosphorylation from 1 of 3 representative experiments (B). Monocytes were pretreated for 30′ in the presence of 3μM RU486 before 16 hours stimulation with DEX (GC) or medium as a control (Co). Subsequently, cells were stimulated to undergo apoptosis by STS (■) or left untreated (□). After 6 hours, the proportion of annexin V–positive cells was measured (C). Data show mean ± SEM (n = 3). In parallel, cells were lysed, and immunoblotting was performed using antiphosphorylated and total ERK1/2 antibodies (D). The figure shows immunoblots from 1 of 3 representative experiments.

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