Figure 4
Hep/D-MKK7 mediates the activation of JNK by cadmium and arsenite. S2 cells were incubated, as indicated, in the absence ("-") of dsRNA, or in the presence of 100 μg per well of dsRNA specific to neomycin resistance gene ("Neo"), or 50 μg per well of dsRNAs specific to D-MKK4, or D-MKK7, or a mixture of both (50 μg each). Four days later, cells were treated with 200 μM CdCl2 or 200 μM NaAsO2 for 2 hours. Expression of D-MKK4 and D-MKK7 mRNAs was detected by RT-PCR using specific primers as indicated (A). "-RT", reactions without Superscript reverse transcriptase; "+RT", reactions with Superscript reverse transcriptase. Cell lysates were immunoprobed with antibodies recognizing P-JNK, P-p38, and P-ERK (B). An experiment representative of 3 repetitions is shown. A quantification of P-JNK (normalized for D-actin) is shown in (C). In this case, the basal levels of P-JNK in each RNAi group (lanes 1, 4, 7, 10, and 13 of Fig. 4B) are represented as "1".