FIGURE 4 from De Novo Purine Metabolism is a Metabolic Vulnerability of Cancers with Low p16 Expression
Year: 2024
Type: other
Abstract: Multiple antifolates induce apoptosis in p16 knockdown cells. A–D, SKMEL28 human melanoma cells were infected with lentivirus expressing a shRNA targeting p16 (shp16). shGFP was used as a control (shCont). A, Cells were treated with the indicated inhibitors (MTX – 0.17 µmol/L; LTX – 0.12 µmol/L) and cytotoxicity was assessed using IncuCyte Cytotox Green reagent. Data from one of three independent experimental replicates are shown (n = 6). Data are mean ± SD. One-way ANOVA at endpoint. ****, P < 0.0001. B, Cells were treated with the indicated inhibitors (MTX – 0.17 µmol/L; LTX – 0.12 µmol/L) for 72 hours, and apoptosis was assessed using Annexin V/PI (propidium iodide) staining by flow cytometry. Data from one of three independent experimental replicates are shown (n = 6). One-way ANOVA of live cells. ****, P < 0.0001. C, Purine metabolite abundance by mass spectrometry. Cells were treated with methotrexate (MTX; 0.17 µmol/L, 72 hours). Data represent one independent experimental replicate (n = 8). Controls are the same data as shown in Fig. 2G. D, Cells were treated with lometrexol (LTX; 0.17 µmol/L) or methotrexate (MTX; 0.17 µmol/L) for 72 hours, and immunofluorescence analysis for γH2AX foci was performed. Data from one of two independent experimental replicates are shown (n = 3). Data are mean ± SD. One-way ANOVA. ****, P < 0.0001; ns = not significant.
Authors: Naveen Kumar Tangudu, Raquel Buj, Hui Wang, Jiefei Wang, Aidan R. Cole, Apoorva Uboveja, Richard Fang, Amandine Amalric, Baixue Yang, Adam Chatoff, Claudia V. Crispim, Peter Sajjakulnukit, Maureen A. Lyons, Kristine Cooper, Nadine Hempel, Costas A. Lyssiotis, Uma R. Chandran, Nathaniel W. Snyder, Katherine M. Aird
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W4396578502