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. 2016 Mar 1;2016:6120509. doi: 10.1155/2016/6120509

Figure 5.

Figure 5

Role of MAPKs p38, JNK, and ERK1/2 in E2-treated bMECs during S. aureus internalization. (a) Participation of MAPKs in S. aureus internalization determined by CFU units. bMECs were incubated (30 min, 1 or 2 h) with pharmacological inhibitors of p38 (5 μM, SB203580), JNK (20 μM, SP600125), or ERK1/2 (2.5 μM, U0126) prior to infection with S. aureus (2 h). Values were determined considering the control (bMECs cultured with the vehicle 1% ethanol) as 100% internalization. (b) The same procedure was used to determine S. aureus internalization by flow cytometry using the LIVE/DEAD BacLight Bacterial viability kit. Fluorescence intensity was estimated from 10,000 events. (c) Phosphorylation was measured in bMECs that were treated with 50 pg/mL E2 and/or challenged with S. aureus by flow cytometry. The phosphorylated MAPK concentrations (U/mL) are represented: (a) pp38, (b) pJNK, and (c) pERK1/2. SB203580: p38 inhibitor. SP600125: JNK1/2 inhibitor. U0126: ERK1/2 inhibitor. According to manufacturer's protocol 300 events were obtained. In (a) and (b) different letters indicate significant changes among treatments (P < 0.05), and in (c) different letters above the bars indicate significant changes among the four treatments within the same MAPK evaluated (P < 0.05). Vehicle: 1% ethanol.