Journal: Frontiers in Immunology

Article Title: Liquid-liquid phase separation-associated gene networks link EMT and ferroptosis to define a PDGFRB-centered prognostic signature in bladder cancer

doi: 10.3389/fimmu.2026.1846096

Figure Lengend Snippet: PDGFRB promotes proliferation, migration, and suppresses ferroptosis in bladder cancer cells. (A) In vitro droplet formation assay of purified recombinant EGFP-PDGFRB-IDR (residues 951–1089) at the indicated concentrations, imaged by confocal microscopy. Scale bar, 2 μm. (B) Western Blot analysis of PDGFRB protein levels in parental T24, shNC (scramble control), and shPDGFRB cells using anti-PDGFRB antibody, with β-actin as loading control. (C) Cell proliferation. CCK-8 assay showing cell viability of shNC and shPDGFRB T24 cells at indicated time points (0, 24, 48, 72, 96 h). (D) Cell migration. Left: Representative images of wound healing assay at 0 and 24 h. Scale bar, 500 μm. Right: Quantification of wound closure rate. (E) Mitochondrial morphology. Transmission electron microscopy images showing mitochondrial morphology in shNC and shPDGFRB T24 cells. Green arrows indicate normal mitochondria with intact cristae; red arrows indicate ferroptotic mitochondria with shrinkage and membrane densification. Scale bar, 0.5 μm. (F) Lipid peroxidation. Left: Ridge plot showing C11-BODIPY fluorescence intensity distribution in shNC and shPDGFRB T24 cells measured by flow cytometry. Right: Quantification of mean fluorescence intensity. (G) MDA levels. Malondialdehyde (MDA) levels in shNC and shPDGFRB T24 cells measured by MDA assay kit. (H) Ferroptosis rescue assay. CCK-8 assay showing cell viability of shNC and shPDGFRB T24 cells with or without Ferrostatin-1 (Fer-1, 2 μM) at indicated time points (0, 24, 48, 72, 96 h). All experiments were performed independently at least three times. For (C, D, F, G) , each independent experiment included two technical replicates (labeled shPDGFRB-1 and shPDGFRB-2) of the same shPDGFRB construct. Data are presented as mean ± s.d. For (B) , significance was calculated by one-way ANOVA. For (C, D, F, G) , significance was calculated by unpaired two-tailed Student's t test. For (H) , significance was calculated by two-way ANOVA. **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Ferrostatin-1 (Fer-1, Targetmol, #347174-05-4) was purchased from Targetmol and dissolved in DMSO.

Techniques: Migration, In Vitro, Tube Formation Assay, Purification, Recombinant, Confocal Microscopy, Western Blot, Control, CCK-8 Assay, Wound Healing Assay, Transmission Assay, Electron Microscopy, Membrane, Fluorescence, Flow Cytometry, Multiple Displacement Amplification, Rescue Assay, Labeling, Construct, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Matrix Remodeling-Associated Protein 8 as a Novel Indicator Contributing to Glioma Immune Response by Regulating Ferroptosis

doi: 10.3389/fimmu.2022.834595

Figure Lengend Snippet: MXRA8 regulates ferroptosis in glioma. (A) The correlation between expression of MXRA8 and ferroptosis-related proteins including NCOA4 and FTH1. (B) NCOA4 and FTH1 were examined by Western blot in U251 and T98G cells transfected with siNC or siMXRA8. (C, D) Fe 2+ and MDA levels were detected in U251 and T98G cells transfected with siNC or siMXRA8. (E) Cell viability was measured in siNC- or siMXRA8-transfected cells after pretreatment with RSL3 (5 μM) and subsequent Fer-1 (20 μM). (F, G) Fe 2+ and MDA levels detected in siNC- or siMXRA8-transfected cells pretreated with RSL3 (5 μM) and Fer-1 (20 μM) for 24 h, successively. (H) After treatment of 5 μM of RSL3 in U251 and T98G cells, Western blot assay were performed to examine the protein expression of MXRA8. * p < 0.05; ** p < 0.01.

Article Snippet: Ras-selective lethal small molecule 3 (RSL3) and ferrostatin-1 (Fer-1) were purchased from APExBIO (B6095 for RSL3, A4371 for Fer-1, Houston, TX, USA).

Techniques: Expressing, Western Blot, Transfection

Journal: Frontiers in Immunology

Article Title: Matrix Remodeling-Associated Protein 8 as a Novel Indicator Contributing to Glioma Immune Response by Regulating Ferroptosis

doi: 10.3389/fimmu.2022.834595

Figure Lengend Snippet: The effects of MXRA8 on M2 macrophage infiltration in vitro by mediating ferroptosis. (A) The correlation coefficient between MXRA8 and classical phenotype markers of macrophages in the GEO datasets. (B) The expression level of MXRA8 and a M2 macrophage phenotype marker (CSF1R) in GBM and normal brain specimens determined by IHC analysis. (C) Infiltration of M2 macrophages in transfected siNC or siMXRA8 cells (U251 and T98G) after pretreatment with RSL3 (5 μM) and subsequent Fer-1 (20 μM). *** p < 0.001.

Article Snippet: Ras-selective lethal small molecule 3 (RSL3) and ferrostatin-1 (Fer-1) were purchased from APExBIO (B6095 for RSL3, A4371 for Fer-1, Houston, TX, USA).

Techniques: In Vitro, Expressing, Marker, Transfection