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. 2007 Apr 3;581(7):1377–1382. doi: 10.1016/j.febslet.2007.02.055

Fig. 1.

Fig. 1

(A) Expression and activation of PKD enzymes in wild-type and PKD1/3−/−DT40 B cells. Cells were treated ± = 25 ng/ml PdBu for 10 min and analyzed by Western blotting of whole cell extracts with the indicated antibodies. PKD1/3−/−DT40 B cells were either continuously maintained in doxycycline (to maintain Flag-PKD3 expression) or were washed out of doxycycline for 5 days (to generate a PKD-null phenotype). (B–D) HSP27 phosphorylation in PKD1/3−/− DT40 B cells. Wild-type, PKD1−/−, PKD3−/− and PKD1/3−/− DT40 B cells were transiently transfected with a Flag-HSP27 expression construct. In Fig. 1D, PKD1/3−/− DT40 B cells were co-transfected with Flag-HSP27 and either GFP, GFP-PKD3 wild-type or GFP-PKD3 K605 N constructs, as indicated. The cells were then left untreated or were stimulated with either 10 μg/ml M 4(BCR) or 25 ng/ml PdBu( P) for 30 min. Flag immunoprecipitates were analysed by SDS–PAGE and Western blotting the indicated antibodies. ∗ = IgL chain.