Figure 7. The C-terminus of DNAJC14 mediates self-interaction.

(A) Self-interaction of DNAJC14. HEK293T cells were cotransfected with pTrip-EGFP-hDNAJC14-NT5 (GFP-NT5) and pV1-hDNAJC14-FL (FL-myc) or the NT5 mutant (NT5-myc) as indicated. Cells were harvested 2 d later and myc-tagged DNAJC14 was immunoprecipitated using anti-myc antibody. Western blots were performed using anti-GFP and anti-myc antibodies as indicated. (B). Self-interaction is mediated by the C-terminus. HEK293T cells were cotransfected as indicated with pTrip-EGFP-hDNAJC14-NT5 (GFP-NT5) and pV1-hDNAJC14-NT5 (NT5-myc, left panels) or the NT5CT1 mutant (NT5CT1-myc, right panels) lacking the C terminal 77 amino acids. Cells were harvested 2 d later and DNAJC14 was immunoprecipitated using anti-GFP or control IgG as indicated. Western blots were performed using anti-GFP and anti-myc antibodies as indicated. For both A and B, arrows indicate the DNAJC14 proteins; migration of size standards (in kDa) is indicated to the left. The asterisk indicates immunoglobulin heavy chain.