Journal: Clinical Cancer Research

Article Title: The Pharmacologic Profile of ART6043, a First-in-Class Clinical DNA Polymerase Theta Polymerase Domain Inhibitor Potentiating PARP Inhibitor Efficacy

doi: 10.1158/1078-0432.CCR-26-0493

Figure Lengend Snippet: ART6043 potentiates PARP inhibition in vitro . A, Representative CTG assay combination matrix for DLD-1 BRCA2 −/− (left) and DLD-1 wild-type (right) cells treated with titrated ART6043 and talazoparib for 14 days. Intensity of blue color indicates the level of synergy. % Control indicates relative viability. B, Representative IncuCyte growth curves for DLD-1 BRCA2 −/− cells treated with ART6043 or talazoparib as monotherapy or in combination at the indicated doses for 14 days. C, As for A but using olaparib instead of talazoparib. D, As for B but using olaparib instead of talazoparib. ART6043 concentrations represent free drug in tissue culture medium.

Article Snippet: Cells were imaged on the IncuCyte Live-Cell Imaging System (Essen Biosciences, RRID:SCR_023147).

Techniques: Inhibition, In Vitro, CTG Assay, Control, Tissue

Journal: Clinical Cancer Research

Article Title: Preclinical Characterization of CLSP-1025, a First-in-Class, Mutation-Specific T-Cell Engager Targeting a Neoantigen Derived from a Common p53 Mutation

doi: 10.1158/1078-0432.CCR-26-1252

Figure Lengend Snippet: CLSP-1025 demonstrates functional activity in various cellular systems including cell cocultures and patient-derived organoids. A, CLSP-1025–mediated cytotoxicity in cocultures of PBMCs and HLA-A*02:01 + –p53 R175H+ tumor cells. Cytotoxicity is reported as the percent tumor-cell death after a 96-hour coculture (mean ± SEM; n = 2). B, HLA-A*02:01 surface expression on p53 R175H+ TYK-nu cells after 72 hours of IFN-γ treatment, measured by geometric mean fluorescence intensity (gMFI) via flow cytometry. Data shown as mean; n = 3 technical replicates. C, CLSP-1025–mediated cytotoxicity in PBMC cocultures with TYK-nu cells with or without IFN-γ pretreatment, shown as percent tumor-cell death (mean ± SEM; n = 3 technical replicates). D–E, CLSP-1025–mediated cytotoxicity and IFN-γ secretion in PBMC cocultures with HLA-A*02:01 + –p53 R175H + adenocarcinoma organoids, measured by percent tumor-cell death and MSD, respectively [mean ± standard deviation (SD); n = 1 PBMC donor and technical duplicates for IFN-γ measurements]. F and G, CLSP-1025–mediated cytotoxicity and IFN-γ secretion in PBMC cocultures with HLA-A*02:01 + –p53 WT adenocarcinoma organoids. H, CLSP-1025–mediated killing of GFP + KLE cells when cocultured with isolated CD4 + , CD8 + , or bulk CD3 + T cells. GFP signal was quantified in real time using Incucyte and plotted as normalized fluorescence units [GFP fluorescence units (GFU) × μm 2 /image]. Data shown as mean ± SD; n = 2 technical replicates.

Article Snippet: The plates were incubated in an Incucyte S3 imager (Sartorius: Incucyte S3 live-cell analysis system, RRID: SCR_023147) for 160 hours and whole-well images captured every 4 hours.

Techniques: Functional Assay, Activity Assay, Patient-Derived Organoids, Expressing, Fluorescence, Flow Cytometry, Organoids, Standard Deviation, Isolation

Journal: Virology Journal

Article Title: Cell culture medium influences RSV G protein glycosylation and recognition by monoclonal antibodies

doi: 10.1186/s12985-026-03318-8

Figure Lengend Snippet: Medium-dependent effects on RSV G protein size, relative protein signals in virus stocks, and viral spread. ( A ) Western blot analysis of soluble G (sG) protein derived from HEK293T cells (top), HEK293T GALE/GALK1 knockout cells (middle), and whole-cell lysates of RSV A-0594-EGFP-infected A549 cells (bottom, 24 hpi), all produced or infected in cell culture medium A, B or C. ( B ) Culture medium-dependent viral spread of RSV A-0594-EGFP in A549 cells infected at an MOI of 1 and cultured in media A, B, or C. GFP integrated fluorescence intensity (GCU × µm²/image; mean ± SD, n = 9 from three independent experiments) was quantified over time using an IncuCyte system (Sartorius). Statistical significance was assessed by two-way ANOVA. ( C ) Virus spread following infection with RSV virus stocks A-C in A549 cells infected at an MOI of 1, with medium replaced by Opti-MEM (medium B) after adsorption. GFP integrated fluorescence intensity (GCU × µm²/image; mean ± SD, n = 9 from three independent experiments) was quantified over time using an IncuCyte system. ( D ) Relative G protein signal normalised to N protein signal in lysed RSV A-0594-EGFP virus stocks A-C, determined by Western blot densitometry (mean ± SD, n = 3). Statistical significance was assessed by one-way ANOVA. ( E ) Relative F protein signal normalised to N protein signal in lysed RSV A-0594-EGFP virus stocks A-C, determined by Western blot densitometry (mean ± SD, n = 3). Statistical significance was assessed by one-way ANOVA. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, **** P ≤ 0.0001. ns, not significant

Article Snippet: Integrated GFP fluorescence (GCU × μm2/image) was quantified over 5 days using an IncuCyte S3 system (Sartorius).

Techniques: Virus, Western Blot, Analysis, Derivative Assay, Knock-Out, Infection, Produced, Cell Culture, Fluorescence, Adsorption

Journal: Virology Journal

Article Title: Cell culture medium influences RSV G protein glycosylation and recognition by monoclonal antibodies

doi: 10.1186/s12985-026-03318-8

Figure Lengend Snippet: Antibody binding and neutralization of recombinant RSV produced in medium A, B or C. ( A ) Quantification of red (therapeutic F-specific antibodies: palivizumab, which binds both the pre-F and post-F conformations, and the pre-F-specific antibodies nirsevimab and clesrovimab) and blue (post-F-specific control antibody MAB8599) fluorescence intensity, normalized to GFP signal, for infected cells cultured in medium A, B or C (mean ± SD, n = 3). Statistical significance was assessed by one-way ANOVA. ( B ) Representative fluorescence images of infected cells stained with palivizumab, nirsevimab and clesrovimab (red) and the post-F control antibody (blue). ( C ) IC50 values of palivizumab, nirsevimab and clesrovimab, against virus produced with medium B or C, determined by 4-PL regression (mean ± SD, n = 3 independent experiments). Statistical significance was assessed using fitted IC50 values. ( D ) Scatter plot showing RT-qPCR Cq values versus TCID50 values for virus stocks A-C. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, **** P ≤ 0.0001. ns, not significant

Article Snippet: Integrated GFP fluorescence (GCU × μm2/image) was quantified over 5 days using an IncuCyte S3 system (Sartorius).

Techniques: Binding Assay, Neutralization, Recombinant, Produced, Control, Fluorescence, Infection, Cell Culture, Staining, Virus, Quantitative RT-PCR