Fig. 8: TNF-α upregulates ILF3 expression by inhibiting TRIM17-mediated ubiquitination. | Cell Death & Disease

Fig. 8: TNF-α upregulates ILF3 expression by inhibiting TRIM17-mediated ubiquitination.

From: ILF3 promotes colorectal cancer cell resistance to ferroptosis by enhancing cysteine uptake and GSH synthesis via stabilizing SLC3A2 mRNA

Fig. 8: TNF-α upregulates ILF3 expression by inhibiting TRIM17-mediated ubiquitination.The alternative text for this image may have been generated using AI.

A Workflow for differential protein identification via pull-down of ILF3 followed by LC/MS analysis. B Protein mass spectrometry identification results. C Venn diagram showing the intersection of 141 differentially expressed proteins identified in SW620 cells treated with TNF-α via LC/MS and 362 E3 ubiquitin ligases retrieved from the UbiNet database. D Co-IP analysis of SW620 cell lysates using an anti-ILF3 antibody or an anti-TRIM17 antibody and control IgG to detect interactions between ILF3 and TRIM17 proteins. E, F HEK293T cells were co-transfected with Flag-TRIM17 or a control vector and Myc-ILF3 for 48 h, followed by IP and IB analyses. G, H SW620 cells treated with TNF-α (40 ng/mL, 24 h) were harvested, and Co-IP was performed using an anti-ILF3 or anti-TRIM17 antibody and control IgG to determine whether TNF-α affects the interaction between ILF3 and TRIM17 proteins. I HEK293T cells were co-transfected with Flag-TRIM17 and Myc-ILF3 for 48 h, treated with TNF-α (40 ng/mL, 24 h) and MG132 (20 μM, 4 h), and subjected to IP and IB analysis. J HEK293T cells were co-transfected with Flag-TRIM17, His-Ub, and Myc-ILF3, treated with TNF-α (40 ng/mL, 24 h) and MG132 (20 μM, 4 h), and subjected to IP and IB analysis to determine whether TNF-α affects ILF3 ubiquitination by TRIM17. K HEK293T cells were transfected with Myc-ILF3 and Flag-TRIM17 together with ubiquitin WT or different ubiquitin mutants. After treatment with TNF-α (40 ng/mL, 24 h) and MG132 (20 μM, 4 h), the cells were collected and subjected to western blot assays. L HEK293T cells were transfected with Myc-ILF3 and Flag-TRIM17 together with ubiquitin K29R mutant. After treatment with TNF-α (40 ng/mL, 24 h) and MG132 (20 μM, 4 h), the cells were collected and subjected to western blot assays.

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