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. 2020 Jan 1;10(3):998–1015. doi: 10.7150/thno.40076

Figure 1.

Figure 1

ZDHHC17-MAP2K4-JNK/p38 Signaling Module Formation in Glioblastoma Multiforme (GBM). (A) The expression of ERK1 (pT202, pY204), ERK2 (pT185, pY187), JNK1/2(pT183, pT185), JNK3 (pT221, pY223) and p38 (pT180, pY182) in GBM is summarized based on the immunohistochemistry results of the Human Protein Atlas. (B) The expression of different DHHCs in glioma is summarized based on the immunohistochemistry results of the Human Protein Atlas. (C) Venn diagram showing the relationship between expression patterns of different DHHCs and activation of ERK1, ERK2, JNK1/2, JNK3, and p38 in GBM. (D) Lysates from HEK293 cells expressing Myc-ZDHHC17 and Flag-MAPKKs were subjected to immunoprecipitation (IP), followed up by immunoblotting (IB) with anti-FLAG antibodies and anti-Myc. IP, immunoprecipitation; IB, immunoblotting. (E) GST pull-down utilizing purified GST-MAP2K4 (or GST-MAP2K7) and FLAG-ZDHHC17-expressing HEK293 cell lysates, followed by IB with an anti-FLAG antibody. (F) ZDHHC17 associates with MAP2K4 via ZDHHC17 ANK rather than PAT activity domain. IP of lysates from HEK293 cells expressing Flag-MAP2K4 and Myc-ZDHHC17 mutants, followed by IB with anti-Flag antibodies and anti-Myc. ZDHHC17 ΔDHHC: ZDHHC17 aa 440-487 deletion; ΔANK: ZDHHC17 aa 51-288 deletion. (G) ZDHHC17 protein recruits MAP2K4 in the Golgi and cytoplasmic vesicles in U118MG cells, in a ZDHHC17 ANK domain-dependent manner. Immunofluorescence of U118MG cells expressing Myc-ZDHHC17 (or -ZDHHC17 ΔDHHC or ΔANK) with anti-Myc (Pink), MAP2K4 (Red), and GM130 (Green) antibodies. Scale bar, 20 µm. (H) ZDHHC17 interacts with MAP2K4 wild-type (wt) and the kinase-inactive mutant (ki). IP of lysates from HEK293 cells expressing Myc-ZDHHC17 and FLAG-MAP2K4wt (or FLAG-MAP2K4ki), followed by IB with anti-FLAG antibodies and anti-Myc. (I) MAP2K4 contributes to the ZDHHC7-mediated JNK1 and p38 phosphorylation. FLAG-MAP2K4ki co-expression in HEK293 cells reduces Myc-ZDHHC17-mediated GFP-JNK1 (or GFP-p38) phosphorylation. (J) MAP2K4 and JNK1 (or p38) are recruited by ZDHHC17 in a signaling module. GFP-JNK1 (p38) and FLAG-MAP2K4 were introduced in HEK293 cells, with or without Myc-ZDHHC17. MAP2K4 presence upon JNK1 (or p38) IP is improved by ZDHHC17 co-expression.