Journal: bioRxiv

Article Title: Can Plitidepsin Be Used as an Antiviral Against RSV?

doi: 10.1101/2025.02.27.640515

Figure Lengend Snippet: Impact of plitidepsin treatment on HRSV replication on HEp-2 cells. (A) Cells were infected for 2 h with rHRSV-mCherry at MOI 0.2 and the medium was then replaced to incubate cells in the presence of serial dilutions of plitidepsin for 48 h (red curve). The viral replication was quantified by measurement of the mCherry fluorescence. In parallel, cell viability upon treatment with plitidepsin was quantified in non-infected HEp-2 cells (black curve). Error bars are standard deviations from duplicates. Data are representative of three experiments. The curves were fitted in Graph Pad 8 software using a four parameters logistic (4PL) regression. Both IC 50 and CC 50 are indicated. (B) Representative images of HEp-2 cell cultures infected with rHRSV-mCherry at MOI 0.2, 48 hours post-infection in the absence or presence of plitidepsin at 12, 4, and 1.4 nM. Nuclei were colored with Hoechst 33432. Scale bars, 250 µm.

Article Snippet: Plitidepsin (MedChemExpress) and Carfilzomib (Cell Signaling #15022) were solubilized in DMSO at 1 mM and 5 mM as stock solutions, respectively.

Techniques: Infection, Fluorescence, Software

Journal: bioRxiv

Article Title: Can Plitidepsin Be Used as an Antiviral Against RSV?

doi: 10.1101/2025.02.27.640515

Figure Lengend Snippet: Plitidepsin effect on a HRSV minigenome. (A) BSRT7/5 cells were transfected with plasmids encoding the N, P, M2–1, and L proteins; the pMT/Luc minigenome, together with pCMV-beta-gal for transfection standardization. Serial dilutions of plitidepsin were added 6 h post-transfection after removing transfection reagents. Viral RNA synthesis was quantified by measuring the luciferase activity after cell lysis 24 h post-transfection. Each luciferase minigenome activity value was normalized based on DMSO treated cells and is the average of three independent experiments performed in triplicate. Error bars represent standard deviations calculated based on three independent experiments made in triplicate. (B) BSRT7 cells were transfected with the complete minigenome system (see above) or with the pEGFP plasmid coding for EGFP. Six hours later, the transfection mix was removed and medium containing plitidepsin at different concentrations was added. Then, 24h post-transfection, cells were lysed and analyzed by Western blotting using anti-N, anti-GFP or anti-alpha tubulin antibodies.

Article Snippet: Plitidepsin (MedChemExpress) and Carfilzomib (Cell Signaling #15022) were solubilized in DMSO at 1 mM and 5 mM as stock solutions, respectively.

Techniques: Transfection, Luciferase, Activity Assay, Lysis, Plasmid Preparation, Western Blot

Journal: bioRxiv

Article Title: Can Plitidepsin Be Used as an Antiviral Against RSV?

doi: 10.1101/2025.02.27.640515

Figure Lengend Snippet: In vitro effect of plitidepsin on HRSV RNA polymerase (L-P complex) activity. Primer extension polymerase assay was performed in the presence of increasing concentration of plitidepsin. Briefly, 200 nM of RSV L/P was incubated with 2 μM of 11 nucleotide length RNA template and 4 nucleotide primer in presence of increasing concentrations of plitidepsin. The reactions were incubated for 30’ and 120 min at 30°C and the reaction products were separated on a 20 % UREA-PAGE gel before gel scanning on a Typhoon imager.

Article Snippet: Plitidepsin (MedChemExpress) and Carfilzomib (Cell Signaling #15022) were solubilized in DMSO at 1 mM and 5 mM as stock solutions, respectively.

Techniques: In Vitro, Activity Assay, Concentration Assay, Incubation

Journal: bioRxiv

Article Title: Can Plitidepsin Be Used as an Antiviral Against RSV?

doi: 10.1101/2025.02.27.640515

Figure Lengend Snippet: Compared effect of plitidepsin on viral or cellular proteins synthesis. HEp-2 cells in 6 well plates were infected with rHRSV-mCherry at MOI = 1. 24 h later, medium was changed for protein labelling with AHA with serial dilutions of plitidepsin. After 4 h, cells were lysed and AHA-labelled proteins were revealed with AFDye 488-DBCO, then immunoprecipitated with anti-N or anti-alpha-tubulin antibodies. Immunoprecipitated proteins or supernatants were resolved by SDS-PAGE (A) and fluorescence in the gels was measured using a Fuji FLA-3000 scanner (B).

Article Snippet: Plitidepsin (MedChemExpress) and Carfilzomib (Cell Signaling #15022) were solubilized in DMSO at 1 mM and 5 mM as stock solutions, respectively.

Techniques: Infection, Immunoprecipitation, SDS Page, Fluorescence

Journal: bioRxiv

Article Title: Can Plitidepsin Be Used as an Antiviral Against RSV?

doi: 10.1101/2025.02.27.640515

Figure Lengend Snippet: Antiviral mechanism of action of plitidepsin is mediated through proteasome-mediated degradation of eEF1A. BSRT7/5 cells were treated with serial dilutions of plitidepsin or DMSO for 18H in the presence or not of the proteasome inhibitor Carfilzomib (500 nM) and the expression of eEF1A or GAPDH were analyzed by WB.

Article Snippet: Plitidepsin (MedChemExpress) and Carfilzomib (Cell Signaling #15022) were solubilized in DMSO at 1 mM and 5 mM as stock solutions, respectively.

Techniques: Expressing

Journal: Cell reports

Article Title: Identification of WNK1 as a therapeutic target to suppress IgH/MYC expression in multiple myeloma.

doi: 10.1016/j.celrep.2024.114211

Figure Lengend Snippet: Figure 6. Combination treatment of WNK463 and other anti-MM compounds exhibits synergistic effects in MM cells (A) Top: cell viability and CI (calculated using the Chou-Talalay method) of JJN3 cells treated with the indicated concentrations of WNK463 and carfilzomib, lenalidomide, or JQ1 for 72 h. Bottom: isobologram plot showing the synergistic effects with each drug combination. (B and C) Cell cycle (B) and apoptosis (C) analysis of JJN3 cells treated with the indicated compounds alone or in combination. Cell cycle and apoptosis analysis were performed after 24 h and 48 h of treatment, respectively. Data are represented as mean ± SD (n = 3). The p values were calculated using one-way ANOVA followed by Dunnett’s multiple-comparisons test. *p < 0.05, **p < 0.01.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins WNK463 MedChemExpress Cat# HY-100626 Carfilzomib Cell Signaling Technology Cat# 15022 Lenalidomide Cell Signaling Technology Cat# 90543 Lenalidomide MedChemExpress Cat# HY-A0003 JQ1 MedChemExpress Cat# HY-13030 Venetoclax MedChemExpress Cat# HY-15531 Erdafitinib MedChemExpress Cat# HY-18708 D-Luciferin, Potassium Salt GoldBio Cat# LUCK-100 DMSO Millipore Sigma Cat# D8418 16% Formaldehyde, Methanol-Free Cell Signaling Technology Cat# 12606 Hydroxypropyl cellulose, M.W.

Techniques: