Journal: Viruses

Article Title: HSV-1 US3 Hijacks Conserved Actin Regulatory Complexes to Drive F-Actin Remodeling

doi: 10.3390/v18070793

Figure Lengend Snippet: Pharmacological validation and inferred model of US3-mediated actin remodeling. ( a ) Schematic of the machine-learning workflow used to prioritize candidate F-actin regulators. Label-independent quantitative mass-spectrometry features were used for model training, whereas curated actin-regulator annotations were used only to define class labels. Prioritized candidates were subsequently selected for biological interpretation and experimental validation. ( b ) Flow-cytometric quantification of F-actin levels in N2A cells infected with WT HSV-1 or ΔUS3 HSV-1 and treated with the formin inhibitor SMIFH2, the Arp2/3 inhibitor CK-666, or both inhibitors. F-actin abundance was measured as the GeoMean MFI of Alexa Fluor 647–phalloidin at 0 and 24 hpi. Inhibition of either pathway partially attenuated the WT-associated reduction in F-actin, whereas combined inhibition prevented a statistically detectable reduction under the tested conditions. Data represent the mean ± SEM from three independent biological experiments. Statistical significance was assessed using two-way ANOVA followed by Šidák’s multiple-comparisons test. ns, not significant; * p < 0.05; **** p < 0.0001. ( c ) Proposed model integrating the US3 IP-MS interactome, machine-learning-prioritized proteins, and experimentally observed F-actin dynamics. Proteins detected in the US3 IP-MS interactome are distinguished from computationally prioritized or inferred proteins. Arrows represent proposed regulatory relationships, many of which may be indirect. The association between US3 and β-catenin was independently validated by reciprocal co-immunoprecipitation . During early infection, US3 is proposed to promote actin-polymerization programs involving Arp2/3- and formin-associated pathways, potentially supporting intracellular transport. At later stages, US3 is proposed to modulate Rho-family GTPase and cofilin signaling, contributing to stress-fiber disassembly and cytoskeletal remodeling associated with viral egress and spread. Myosin-associated motor complexes may contribute to cytoskeletal organization during both phases.

Article Snippet: Specifically, the formin inhibitor SMIFH2 (MedChemExpress, Monmouth Junction, NJ, USA; 20 μM) and the Arp2/3 complex inhibitor CK-666 (MedChemExpress, Monmouth Junction, NJ, USA;100 μM) were used.

Techniques: Biomarker Discovery, Mass Spectrometry, Infection, Inhibition, Protein-Protein interactions, Immunoprecipitation

Journal: Cell Death & Disease

Article Title: UBC9-mediated SUMOylation of CORO1C drives lung adenocarcinoma progression via Arp2/3-dependent cytoskeletal remodeling

doi: 10.1038/s41419-026-08653-w

Figure Lengend Snippet: A549 cells reconstituted with WT-CORO1C or 3KR mutant were treated with DMSO or 100 μM CK-666 for 30 h. A Representative images of phalloidin-stained F-actin (red) in the indicated cell lines treated with DMSO or CK-666. Nuclei are stained with DAPI (blue). Scale bar, 20 μm. CCK-8 proliferation assay ( B ) and clonogenic assay ( C ) of the indicated cell lines treated with DMSO or CK-666. D Quantification of colony numbers from ( C ). E Representative images of Transwell migration (upper) and Matrigel invasion (lower) assays after CK-666 treatment. Scale bar, 50 μm. ( F , G ) Quantitative analysis of migrated and invaded cells from ( E ). H Tumor growth curves of subcutaneous xenografts in mice with or without intraperitoneal injection of CK-666 (20 mg/kg) once a week ( n = 5). I Photographs of dissected tumors from each group at the endpoint ( n = 5). J Final tumor weights in each group. In vivo bioluminescence imaging of orthotopic tumors derived from luciferase-expressing A549 CORO1C WT or 3KR cells in mice with or without intraperitoneal CK-666 treatment. K Representative bioluminescence images. L Quantitative analysis of in vivo radiance signals in ( K ). Data are expressed as means ± SD. M Representative H&E-stained lung sections of metastatic nodules from mice with or without intraperitoneal injection of CK-666 (20 mg/kg) once a week. Arrows indicate metastatic foci. N Quantitative analysis of metastatic foci from ( M ). Quantitative data are expressed as means ± SD.

Article Snippet: For pharmacological inhibition of the Arp2/3 complex, cells were treated with CK-666 (MCE, #HY-16926) at a final concentration of 100 μM.

Techniques: Mutagenesis, Staining, CCK-8 Assay, Proliferation Assay, Clonogenic Assay, Migration, Injection, In Vivo, Imaging, Derivative Assay, Luciferase, Expressing