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. 2004 Oct 26;383(Pt 3):447–455. doi: 10.1042/BJ20040498

Figure 5. Phosphorylation of JNK and p38 upon sorbitol treatment is enhanced by LDP-3.

Figure 5

COS-7 cells were co-transfected with 0.5 μg of pSRα-HA-JNK1 (A), pMT3-HA-p38α (B) or pSRα-HA-ERK2 (C and D) together with 1.5 μg of pFLAG-CMV, pFLAG-LDP-3, pFLAG-VHR, pFLAG-MKP-2 or pFLAG-MKP-5. After 36 h, transfected cells were stimulated with 0.4 M sorbitol for 30 min (A, B and D) or 50 ng/ml EGF for 15 min (C). Phosphorylated HA–JNK1, HA–p38α and HA–ERK2 were detected by anti-ACTIVE JNK, anti-phospho-p38α and anti-phospho-ERK antibodies respectively. The expression levels of HA–MAPKs (HA–JNK1, HA–p38α and HA–ERK2) and FLAG-tagged phosphatases (FLAG–LDP-3, FLAG–VHR, FLAG–MKP-2 and FLAG–MKP-5) were assessed by immunoblot using anti-HA (12CA5) and anti-FLAG M2 antibodies respectively.