Journal: Cell Death Discovery

Article Title: IL‑1 receptor antagonism attenuates renal fibrosis via RNF182‑driven MFN2 destabilization and mitochondrial dysfunction

doi: 10.1038/s41420-025-02929-4

Figure Lengend Snippet: In the context of kidney injury, pro-inflammatory stimuli upregulate the expression of the Interleukin-1 Receptor (IL-1R) and its downstream effector, TGF-β1, in renal tubular epithelial cells. This leads to increased expression of the E3 ubiquitin ligase RNF182. RNF182, in turn, forms a complex with the mitochondrial fusion protein Mitofusin 2 (MFN2), targeting it for ubiquitination and subsequent proteasomal degradation. The depletion of MFN2 results in mitochondrial dysfunction, characterized by excessive reactive oxygen species (ROS) production and reduced ATP synthesis, which drives the progression of renal fibrosis. Therapeutic administration of recombinant human IL-1 receptor antagonist (rhIL-1Ra) blocks IL-1R signaling. This intervention suppresses the downstream upregulation of RNF182, thereby reducing MFN2 ubiquitination and degradation. The stabilization of MFN2 protein preserves mitochondrial function, mitigates oxidative stress, and restores cellular energy production, ultimately exerting a potent anti-fibrotic effect and protecting kidney tissue.

Article Snippet: Cells grown on coverslips were fixed with 4% PFA for 15 min, permeabilized with 0.2% Triton X-100 in PBS for 10 min, and blocked with 5% BSA in PBS for 1 h. Cells were incubated with primary antibodies against IL-1R (R&D Systems, Cat# AF269, 1:100 dilution) overnight at 4 °C.

Techniques: Expressing, Ubiquitin Proteomics, Recombinant

Journal: Biofabrication

Article Title: A microphysiological assay for studying T-cell chemotaxis, trafficking and tumor killing.

doi: 10.1088/1758-5090/ad847f

Figure Lengend Snippet: Figure 2. 2-lane assay arrangement for T-cell chemotaxis (a) Changes made to the pre-gel additional sequence to achieve aligned collagen fibers (ECM: 2 mg ml−1 collagen mixed with 10% matrigel GFR) (b) three different assay arrangements (i), (ii) and (iii) to understand the best arrangement for robust activated CD3+ T-cell chemotaxis response to CXCL12 gradient. T-cell migration distances were analyzed and quantified at 120 h after experimental initiation (a representative image). Blue arrow indicates the direction of cell motion and the x = 0 chemotaxis distance. (c) Final assay arrangement to study immune cell chemotaxis against chemotactic stimuli. Direction of the gradient and resulting cell chemotaxis is shown. Scale bar = 1000 µm (linear mixed effects model corrected for multiple comparisons, ∗∗∗∗p-value < 0.0001).

Article Snippet: Recombinant IL2 (Cat No: 130- 097-748), CXCL12 (Cat No: 350-NS) and CXCL10 (Cat No: 266-IP) were purchased from R&D systems (Minneapolis, MN).

Techniques: Chemotaxis Assay, Sequencing, Migration

Journal: Biofabrication

Article Title: A microphysiological assay for studying T-cell chemotaxis, trafficking and tumor killing.

doi: 10.1088/1758-5090/ad847f

Figure Lengend Snippet: Figure 3. Activated CD3+ T cell responses to CXCL12 and CXCL10 gradient. Activated CD3+ T-cell chemotaxis response against (a) CXCL12 and (b) CXCL10 at 96 h within end-to-end ECM filled 2-lane plates. Numbers of T-cells under migration and T-cell migration length is shown (median T-cell migration distances indicated by black arrow). Inserts show representative images of 0 nM and 300 nM CXCL12 and CXCL10 split across two fields of images (taken at 4X magnification). (one-way ANOVA corrected for multiple comparisons, ∗p-value < 0.05, ∗∗p-value < 0.01, ∗∗∗∗p-value < 0.0001).

Article Snippet: Recombinant IL2 (Cat No: 130- 097-748), CXCL12 (Cat No: 350-NS) and CXCL10 (Cat No: 266-IP) were purchased from R&D systems (Minneapolis, MN).

Techniques: Chemotaxis Assay, Migration

Journal: Cell Reports Methods

Article Title: Tongue orthotopic xenografts to study fusion-negative rhabdomyosarcoma invasion and metastasis in live animals

doi: 10.1016/j.crmeth.2024.100802

Figure Lengend Snippet:

Article Snippet: Lymphatics were visualized by injecting 0.4ug of Mouse LYVE-1 Alexa Fluor 594-conjugated Antibody (20ul of concentration 20ug/ml, diluted 1:10 in normal saline; cat# FAB2125T; R&D systems) into the tongue in a region near the tumor 20 min before imaging.

Techniques: Recombinant, Virus, Membrane, Injection, Electron Microscopy, Software, Imaging