Journal: Viruses

Article Title: Establishment of a Dual-Reporter Minigenome System for Respiratory Syncytial Virus

doi: 10.3390/v18030304

Figure Lengend Snippet: Mini-NLuc-sfGFP for small-molecule evaluation. ( A ) The experimental protocol involved transfecting the Mini-NLuc-sfGFP minigenome into BSR-T7/5 cells, followed by treatment with a range of small molecules. The inhibitory effects were initially assessed by quantifying the fluorescence intensity or NLuc activity, which led to the selection of promising candidates for further investigation. Created in BioRender. Wu, C. (2026) https://BioRender.com/8qct7m8 . ( B ) AVG-233 reduced the BSR-T7/5 cell viability in a concentration-dependent manner (OD 450 ). ( C ) Similarly, RSV L-protein-IN-4 reduced the BSR-T7/5 cell viability in a concentration-dependent manner (OD 450 ). ( D ) AVG-233 demonstrated a dose-dependent inhibition of Mini-NLuc-sfGFP reporter gene expression, as indicated by the luciferase activity. ( E ) Similarly, RSV L-protein-IN-4 exhibited the dose-dependent inhibition of the Mini-NLuc-sfGFP reporter gene expression, also measured using the luciferase activity. Data are presented as the mean ± SD of three independent experiments, each performed in triplicate.

Article Snippet: Small-molecule inhibitors of RSV L polymerase, namely, AVG-233 (CAS no. 2151937-80-1) and RSV L-protein-IN-4 (CAS no. 851657-60-8), both sourced from Topscience Co. Ltd., China, were diluted in DMSO to create stock solutions at various concentrations.

Techniques: Fluorescence, Activity Assay, Selection, Concentration Assay, Inhibition, Gene Expression, Luciferase

Journal: Viruses

Article Title: Establishment of a Dual-Reporter Minigenome System for Respiratory Syncytial Virus

doi: 10.3390/v18030304

Figure Lengend Snippet: Validation of small-molecule inhibitory effects assessed by Mini-NLuc-sfGFP and confirmed in RSV A2. RSV A2 infection was conducted at a multiplicity of infection (MOI) of 0.1, utilizing a primary antibody of mouse anti-RSV F and a secondary antibody of goat anti-mouse FITC conjugate. ( A ) Immunofluorescence imaging showed that AVG-233 inhibited viral infection at 0, 0.01, 0.05, 0.25, 0.5, 1, 2, and 3 μM. ( B ) Viral genome copy numbers via RT-qPCR at equivalent concentrations. ( C ) Immunofluorescence imaging showed that RSV L-protein-IN-4 inhibited viral infection at 0, 0.01, 0.05, 0.25, 0.5, 1, 2, and 3 μM. ( D ) Viral genome copy numbers via RT-qPCR at equivalent concentrations. Scale bar: 400 μm. Data are presented as the mean ± SD of three independent experiments, each performed in triplicate.

Article Snippet: Small-molecule inhibitors of RSV L polymerase, namely, AVG-233 (CAS no. 2151937-80-1) and RSV L-protein-IN-4 (CAS no. 851657-60-8), both sourced from Topscience Co. Ltd., China, were diluted in DMSO to create stock solutions at various concentrations.

Techniques: Biomarker Discovery, Infection, Immunofluorescence, Imaging, Quantitative RT-PCR

Journal: EMBO Molecular Medicine

Article Title: Micrometastasis-derived models enable drug testing for early-stage, high-risk melanoma patients

doi: 10.1038/s44321-025-00339-8

Figure Lengend Snippet: ( A ) Upper graph: Vemurafinib treatment of Mel-DCC CLs. Cells were incubated with doses ranging from 0.025 µM to 5 µM Vemurafenib for 5 days. Cell viability is shown for BRAF wt Mel-DCC-07 (red, n = 4), BRAF V600K-mutated Mel-DCC-13 (gray, n = 4), and BRAF V600E-mutated Mel-DCC-02 (black, n = 5). Lower graph: Binimetinib treatment of Mel-DCC CLs. Cells were incubated with Binimetinib at doses ranging from 0.001 µM to 1 µM for 5 days. Cell viability is shown for NRAS Q61R-mutated Mel-DCC-04 (red, n = 4), NRAS T58I-mutated Mel-DCC-07 (gray, n = 6), and NRAS Q61K-mutated Mel-DCC-01 (black, n = 4). Each dot represents the mean value ± SD of biological replicates. ( B ) Generation of a Vemurafenib-resistant BRAF-mutated melanoma cell line (Mel-DCC-11-R). Resistance was generated through stepwise exposure to increasing concentrations of Vemurafenib over the indicated timeframe. Sensitivity of Mel-DCC-11 (black, n = 3) vs. Mel-DCC-11-R (red, n = 5) to Vemurafenib is shown. Each dot represents the mean value ± SD of biological replicates. ( C ) Outcome of experimental drug testing with 315 anti-cancer drugs on BRAF V600E-mutated Mel-DCC-11 and Vemurafenib-resistant Mel-DCC-11-R, alone or in combination with 8 µM Vemurafenib (Mel-DCC-11-R + V). The number of drugs that reduce cell viability to less than 80% is indicated. ( D ) Heatmap showing the drug-induced reduction of the viability in the Vemurafenib-restistant CL, screened in the presence (Mel-DCC-11-R + V) or absence (Mel-DCC-11-R) of Vemurafenib, alongside the parental Vemurafenib-sensitive Mel-DCC-11, screened without Vemurafenib. The mean viability of two biological replicates is shown. .

Article Snippet: Anti-Cancer Approved Drug Library (315 compounds) , TargetMol , Cat# L2110.

Techniques: Incubation, Generated