Xenbase Image ID: 155104

Figure 6. Interaction of DVL1 and DVL2 with PKD1 in MEFs and transiently transfecte HEK293T cells(a) Wild type MEF cell lysates were incubated with mouse IgG1 control antibody or mouse monoclonal α-PKD1 (E4). Immunocomplexes were immunoblotted with rabbit polyclonal α-DVL1 (upper left panel), α-DVL2 (middle left panel), or mouse monoclonal α-PKD1 (7e12, lower left panel). Expression levels of DVL1, DVL2, or PKD1 in lysates incubated with mouse IgG1 control or α-PKD1 (E4) are shown in right panels (input). Experiments were successfully repeated 3 times. (b) Cell lysates from Pkd1−/− or Pkd2−/− MEFs were incubated with mouse IgG1 control antibody or mouse monoclonal α-PKD1 (E4) and DVL2 was detected in the immunocomplexes, as described in (a). Experiments were successfully repeated twice. (c) HEK293T cells were transiently transfected with indicated plasmids and PKD1 or TRPP2 was pulled down with α-HA. Flag-tagged wild type or mutant DVL2 was detected using α-Flag. DIX (DIshevelled and aXin), PDZ, or DEP (Dishevelled, Egl-10 and Plecstrin) domains are shown red in DVL2 diagram. Position of E499G mutation and PY motif are shown by a black arrow head. Experiments were successfully repeated 3 times. (d) Time course, step currents and I-V curves in zero intracellular Ca2+ of wild type MEFs transiently transfected with a DVL2-specific siRNA (n=18 cells pooled from 5 independent experiments) or a mixture of DVL1 and DVL2 siRNAs (n=13 cells pooled from 4 independent experiments). Statistical tests were performed using paired Student's t-test, **P<0.01. Data are shown as mean ± SEM. (e) Knockdown efficiency of DVL1 and DVL2 siRNAs. Wild type MEFs were transiently transfected with indicated siRNAs and cell lysates were immunoblotted with α-DVL1, α-DVL2, or α-β-actin. Experiment was done once. Unprocessed original scans of blots are shown in Supplementary Fig. 8). Image published in: Kim S et al. (2016) Image downloaded from an Open Access article in PubMed Central. Image reproduced on Xenbase with permission of the publisher and the copyright holder. Larger Image Printer Friendly View

Return to previous page