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Activation of SAPKs/JNKs and p38s In Vitro
The stress-activated protein kinases (SAPKs)/c-Jun N2 -terminal kinases (JNKs) and the p38 mitogen-activated protein kinases (MAPKs) are two MAPK groups activated predominantly by stresses and proinflammatory stimuli. Table 1 lists some of the stimuli known to recruit both the SAPKs/JNKs and p38s. In general, the same stimuli that activate the SAPKs/JNKs will also activate the p38s. One exception is ischemia-reperfusion. In this instance, the SAPKs/JNKs are activated exclusively during reperfusion, while the p38s are activated during ischemia and remain active during reperfusion (1 ). Table 1 Representative Stimuli That Activate SAPK/JNK and p38 In Vivo a
| Stimulus (ref.) | Target kinase(s) | Sensitive cell lines | Amount used | Comments |
|---|---|---|---|---|
| Anisomycin (6 ) | SAPK/JNK, p38 | All cells | 1–10 μg/mL, 1–60 min | |
| UV-C (12 ) | SAPK/JNK, p38 | All adherent cells | 40 J/m2 , 20 s-1 | Drain medium from cells, and wash with PBS before exposure. |
| γ Radiation (13 ) | SAPK/JNK, p38 | All cells (see comments) | 10–20 gy, 137Cs source at 0.76 gy/min | This is controversial and has not been replicated in some laboratories. After treatment, wait between 20 min and 4 h before harvest. |
| cis platinum (CDDP) or cytosine arabinoside (ara-C) (13 ) | SAPK/JNK, p38 | All cells | 50–300 μM , 2–6 h | |
| Methyl methane sulfonate (MMS) (13 ) | SAPK/JNK, p38 | All cells | 1 mM , 3 h | |
| Oxidant stress (H2 O2 )(9 ) | SAPK/JNK, p38 | All cells | 2–20 mM , 2–60 min | |
| TNF (6 ) | SAPK/JNK, p38 | HepG2, CCD18-CO, HEK293, COS, U937, HL-60, Jurkat, L929, HeLa, primary thymocytes or hepatocytes | 2–100 ng/mL, 1–60 min | Try to pick a cell type in which TNF is known to induce a biologic effect. |
| IL-1 (6 ) | SAPK/JNK, p38 | EL-4, HepG2, Jurkat, HL-60, KB | 2–100 ng/mL, 1–60 min | Try to pick a cell type in which IL-is known to induce a biologic effect. |
| Endothelin (14 , 15 ) | SAPK/JNK, p38 | Endothelial, mesangial, pulmonary fibroblast, vascular smooth muscle, cardiomyocyte | 10–200 nM , 2–60 min | Try to pick a cell type in which endothelin is known to induce a biologic effect. |
| Angiotensin II (14 ) | SAPK/JNK | Endothelial, mesangial, pulmonary fibroblast, vascular smooth muscle, cardiomyocyte, | 20–500 nM , 2–60 min | Try to pick a cell type in which the type II antiotensin II receptor is expressed and in which angiotensin II is known to induce a biologic effect. |
| Ischemia/reperfusion (16 ) | SAPK/JNK, p38 | Kidney, heart, brain | 40 min ischemia, 2–120 min reperfusion | SAPK/JNK is activated only during reperfusion. |
a This table is meant to be a guide for stimuli that recruit SAPK/JNK and p38. The lists of stimuli and cells used are in no way inclusive. Theconditions for stimulation are also meant to serve as a guide. We strongly recommend that time courses and dose responses be performed whenever possible.
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