Journal: Journal of Translational Medicine

Article Title: Polystyrene microspheres could inhibit the ferroptosis of prostate cancer cells via USP39/IGF2BP3/MAPK4 axis

doi: 10.1186/s12967-025-06868-7

Figure Lengend Snippet: PS microspheres promoted the progression of PC cells by regulating USP39. ( A ) A heatmap illustrating the differentially expressed genes in PC cells before and after treatment with PS microspheres. ( B ) A volcano plot depicting the differentially expressed mRNAs. ( C ) PCR analysis showed the expression levels of MAGEA1, SLC1A5, and USP39 in the PS-treated and control groups. ( D ) Western blot analysis demonstrated the expression levels of MAGEA1, SLC1A5, and USP39 in PC cells following PS treatment. CCK-8 assays showed that the proliferative capacity of PC3 ( E ) and DU-145 ( F ) cells was enhanced by PS treatment, an effect that was reversed upon USP39 silencing. ( G ) The colony formation assay revealed that the proliferation capacity of PC cells increased after PS treatment, and this effect was reversed following USP39 silencing. ( H ) The Transwell assay demonstrated the migration and invasion capabilities of PC cells across different groups. ( I ) Western blot analysis revealed the expression levels of markers related to proliferation and migration in the different groups of PC cells. * P < 0.05, ** P < 0.005, *** P < 0.001

Article Snippet: Following permeabilization, the cells were treated with a Tris-buffered saline solution supplemented with 5% BSA for 30 min. Incubating the cells overnight at low temperature (4 °C) preserves their viability and ensures sufficient time for antibody-antigen interactions, especially IGF2BP3 (Proteintech, China) or USP39 (Proteintech, China).

Techniques: Expressing, Control, Western Blot, CCK-8 Assay, Colony Assay, Transwell Assay, Migration

Journal: Journal of Translational Medicine

Article Title: Polystyrene microspheres could inhibit the ferroptosis of prostate cancer cells via USP39/IGF2BP3/MAPK4 axis

doi: 10.1186/s12967-025-06868-7

Figure Lengend Snippet: USP39 can bind to IGF2BP3 and promote its ubiquitination. ( A ) Flowchart of the IP-Mass spectrometry process. ( B ) Proteins identified from mass spectrometry analysis. ( C ) Mass spectrum data for IGF2BP3. ( D ) Illustration of the molecular docking simulation between USP39 and IGF2BP3. ( E ) Immunofluorescence (IF) assay showing the subcellular colocalization of USP39 and IGF2BP3 in PC cells. ( F ) Co-immunoprecipitation (Co-IP) assay demonstrating the interaction between USP39 and IGF2BP3. ( G ) Western blot analysis showing the expression level of IGF2BP3 at different concentrations of PS microspheres. ( H ) Western blot analysis showing the expression level of IGF2BP3 in PC cells at various time points after CHX treatment. ( I ) Western blot analysis showing the expression level of IGF2BP3 in PC cells at different time points after MG-132 treatment. ( J ) Western blot analysis showing the expression level of IGF2BP3 in PC cells at different time points after CQ treatment. ( K ) The level of IGF2BP3 ubiquitination in PC cells, as detected by western blot assay. * P < 0.05, ** P < 0.005, *** P < 0.001

Article Snippet: Following permeabilization, the cells were treated with a Tris-buffered saline solution supplemented with 5% BSA for 30 min. Incubating the cells overnight at low temperature (4 °C) preserves their viability and ensures sufficient time for antibody-antigen interactions, especially IGF2BP3 (Proteintech, China) or USP39 (Proteintech, China).

Techniques: Ubiquitin Proteomics, Mass Spectrometry, Immunofluorescence, Co-Immunoprecipitation Assay, Western Blot, Expressing