Journal: Signal Transduction and Targeted Therapy

Article Title: Inhibin beta A drives colorectal cancer progression through macrophage M2 polarization and mitochondria-dependent ferroptosis suppression

doi: 10.1038/s41392-025-02518-y

Figure Lengend Snippet: INHBA acts as a scaffold protein to prevent the TRIM21-driven ubiquitination and degradation of SLC25A10. a Western blot of SLC25A10 in CRC cells with INHBA silenced or overexpressed after MG132 treatment (20 μM, 4 h). b CHX chase (50 μg/ml, TargetMol, USA) of INHBA-knockdown CRC cells; SLC25A10 abundance determined by Western blot at 0, 3, 6, 9 h. c IP was performed using anti-Flag to detect INHBA-interacting proteins. d IP was performed using anti-HA to identify TRIM21-interacting proteins. e IP was were performed using anti-HA to detect SLC25A10-interacting proteins. f IP was performed using anti-HA to identify TRIM21-interacting proteins. g Enrichment and detection of SLC25A10-interacting proteins: CRC cells were transfected with HA-TRIM21 together with Myc-SLC25A10, Flag-INHBA, or an empty vector. 48 h post-transfection, cells were exposed to MG132 (20 μM, 4 h). Myc-SLC25A10 was immunoprecipitated with anti-Myc beads, and immunoprecipitates were detected via Western blot with anti-HA. h Detection of SLC25A10 ubiquitination levels: CRC cells were transfected with Myc-SLC25A10 and HA-Ub together with Flag-INHBA, His-TRIM21, or an empty vector. 48 h post-transfection, cells were exposed to MG132 (20 μM, 4 h). Myc-SLC25A10 was immunoprecipitated with anti-Myc beads, and ubiquitination was detected by anti-Ub Western blot. i Detection of SLC25A10 ubiquitination levels (K48 and K63): CRC cells were transfected with Myc-SLC25A10 together with HA-Ub-K63, HA-Ub-K48, Flag-INHBA, or empty vector. 48 hours post-transfection, cells were exposed to MG132 (20 μM, 4 h). Myc-SLC25A10 was immunoprecipitated with anti-Myc beads, and ubiquitination was detected by anti-Ub Western blot. j Detection of SLC25A10 ubiquitination levels (K48 linkage): HA-Ub-K48 and Myc-SLC25A10 were co-transfected into CRC cells together with Flag-INHBA, His-TRIM21, or empty vector. After 48 h, cells were exposed to MG132 (20 μM, 4 h). Myc-SLC25A10 was immunoprecipitated with anti-Myc beads, and ubiquitination was detected by anti-Ub Western blot

Article Snippet: Fig. 6 INHBA acts as a scaffold protein to prevent the TRIM21-driven ubiquitination and degradation of SLC25A10. a Western blot of SLC25A10 in CRC cells with INHBA silenced or overexpressed after MG132 treatment (20 μM, 4 h). b CHX chase (50 μg/ml, TargetMol, USA) of INHBA-knockdown CRC cells; SLC25A10 abundance determined by Western blot at 0, 3, 6, 9 h. c IP was performed using anti-Flag to detect INHBA-interacting proteins. d IP was performed using anti-HA to identify TRIM21-interacting proteins. e IP was were performed using anti-HA to detect SLC25A10-interacting proteins. f IP was performed using anti-HA to identify TRIM21-interacting proteins. g Enrichment and detection of SLC25A10-interacting proteins: CRC cells were transfected with HA-TRIM21 together with Myc-SLC25A10, Flag-INHBA, or an empty vector.

Techniques: Ubiquitin Proteomics, Western Blot, Knockdown, Transfection, Plasmid Preparation, Immunoprecipitation

Journal: Cancer Cell International

Article Title: INHBA promotes chemoresistance in pancreatic cancer by enhancing CTPS1 stability and mediating pyrimidine metabolism

doi: 10.1186/s12935-025-03984-8

Figure Lengend Snippet: INHBA interacts with CTPS1. A–C . Silver staining (a) and mass spectrometry (b, c) revealed INHBA-associated proteins in INHBA-overexpressing PANC-1 PC cells. D. Venn diagram showing protein intersection from the pyrimidine metabolism pathway and immunoprecipitation-mass spectrometry analysis. E. Co-IP assays confirmed the interaction between INHBA and CTPS1. F . qRT-PCR detection of the correlation between the transcriptional expression levels of INHBA and CTPS1. G. Immunofluorescence analysis of INHBA and CTPS1 colocalization in tissues and cells (scale bar = 10 μm) H. Western blotting revealed reduced CTPS1 expression following INHBA knockdown. I . MG132 (20 µM) treatment reduced CTPS1 protein degradation in INHBA-knockdown cells compared to that in untreated cells. J , K. With prolonged CHX (20 µM) treatment, CTPS1 degradation accelerated in the INHBA-knockdown group compared to that in the control group, whereas it gradually slowed in the INHBA-overexpression group

Article Snippet: The chemical reagents used in the experiments included CHX (MCE, USA), MG132 (MCE, USA), and DAPI (Solarbio, China).

Techniques: Silver Staining, Mass Spectrometry, Immunoprecipitation, Co-Immunoprecipitation Assay, Quantitative RT-PCR, Expressing, Immunofluorescence, Western Blot, Knockdown, Control, Over Expression