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. 2008 Dec 31;40(6):596–606. doi: 10.3858/emm.2008.40.6.596

Figure 4.

Figure 4

Effect of PKC inhibitors on TonEBP mobility shift under hypertonic conditions. (A) HSF1 mobility shift and translocation to the nucleus by the treatment of heat shock (HS). NIH-3T3 cells were treated with 42℃ heat shock for 30 min and recovered at 37℃ for the indicated time periods. Then, the whole cell lysates, cytosolic fraction, and nuclear fraction were prepared as described in the Materials and Methods section. Proteins were resolved via SDS-PAGE on 8% low-bias gels and blotted with anti-HSF1 and anti-lamin A antibodies. (B) HSF1 status under hypertonic conditions. NIH-3T3 cells were exposed to hypertonic conditions for the indicated time periods. Prepared whole cell lysates, cytosolic fraction, and nucleus fractions were analyzed via immunoblot assay. (C) Effect of PKC inhibitors on the mobility shift of TonEBP. Cells were exposed to hypertonic conditions either for the indicated time periods without any pretreatment (left panel) or for 2 h with pretreatment of 25 µM GF109203X or 10 µM Gö6976 (right panel). Cell lysates were prepared and analyzed via immunoblot assay. (D) Effect of PKC inhibitors on TonEBP translocation. Cells were maintained in hypertonic media for either the indicated time periods without inhibitor treatment (upper panel) or for 2 h with pretreatment of 25 µM GF109203X (middle panel) and 10 µM Gö6976 (lower panel). Cell lysates were prepared and analyzed via immunoblot assay for the dectection of TonEBP translocation.