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MZT2A promotes NEMO ubiquitination by inhibiting the deubiquitinating function of OTULIN. A , the level of NEMO ubiquitination in A549 cells overexpressing MZT2A or under hypoxia by Co-IP analysis; n = 3, ∗ p < 0.05 versus control. B , the interaction between MZT2A and OTULIN in A549 cells by Co-IP analysis. C , GST-pull-down result showed that MZT2A could directly interact with OTULIN. D , the protein levels of MZT2A and OTULIN in A549 cells transfected with MZT2A cDNA, OTULIN-specific siRNA, or both MZT2A cDNA and OTULIN-specific siRNA by Western blot analysis. n = 3, ∗ p < 0.05, ∗∗ p < 0.01 versus control. E , the level of NEMO ubiquitination by Co-IP analysis and the level of IKK phosphorylation by Western blot analysis in A549 cells transfected with MZT2A-specific siRNA, OTULIN-specific siRNA, or OTULIN-specific siRNA and MZT2A cDNA. cDNA, complementary DNA; Co-IP, coimmunoprecipitation; GST, glutathione- S -transferase; MZT2A, mitotic spindle–organizing protein 2A; NEMO, NF-κB essential modulator; OTULIN, OTU deubiquitinase with linear linkage specificity.

Journal: The Journal of Biological Chemistry

Article Title: Mitotic spindle–organizing protein 2A (MZT2A) promotes cisplatin resistance through NEMO ubiquitination and NF-κB activation in lung adenocarcinoma

doi: 10.1016/j.jbc.2026.111330

Figure Lengend Snippet: MZT2A promotes NEMO ubiquitination by inhibiting the deubiquitinating function of OTULIN. A , the level of NEMO ubiquitination in A549 cells overexpressing MZT2A or under hypoxia by Co-IP analysis; n = 3, ∗ p < 0.05 versus control. B , the interaction between MZT2A and OTULIN in A549 cells by Co-IP analysis. C , GST-pull-down result showed that MZT2A could directly interact with OTULIN. D , the protein levels of MZT2A and OTULIN in A549 cells transfected with MZT2A cDNA, OTULIN-specific siRNA, or both MZT2A cDNA and OTULIN-specific siRNA by Western blot analysis. n = 3, ∗ p < 0.05, ∗∗ p < 0.01 versus control. E , the level of NEMO ubiquitination by Co-IP analysis and the level of IKK phosphorylation by Western blot analysis in A549 cells transfected with MZT2A-specific siRNA, OTULIN-specific siRNA, or OTULIN-specific siRNA and MZT2A cDNA. cDNA, complementary DNA; Co-IP, coimmunoprecipitation; GST, glutathione- S -transferase; MZT2A, mitotic spindle–organizing protein 2A; NEMO, NF-κB essential modulator; OTULIN, OTU deubiquitinase with linear linkage specificity.

Article Snippet: Plasmids carrying the human MZT2A cDNA (#RC219963), NEMO cDNA (#RC201743), and their corresponding vector controls were obtained from OriGene.

Techniques: Ubiquitin Proteomics, Co-Immunoprecipitation Assay, Control, Transfection, Western Blot, Phospho-proteomics