Fig. 3.

GILZ knockdown increased basal phosphorylation of ERK1/2 and JNK, and p65 NF-κB nuclear translocation. A: Control or GILZ-silenced human adipocytes were treated with TNFα (3 ng/ml) for 1 h, and ERK1/2, JNK, and p65 NF-κB phosphorylation and MKP-1 expression levels were determined with Western blotting. HSP90 was used as a loading control. B: Quantification of ERK1/2 phosphorylation relative to total ERK1/2. C: Quantification of JNK phosphorylation relative to total JNK. ^##P < 0.01 compared with control siRNA, n = 5. D: After treating with TNFα (3 ng/ml, 1 h), nuclear extracts (NE) and cytoplasmic extracts (CE) were prepared and used for measurement of p65 NF-κB. α-Tubulin and RNA POLII were used as markers of cytoplasmic and nuclear extracts, respectively. Representative blots of two (for MKP-1) or three experiments are shown. ctrl, control.