Additional file 1 of Modulating tumour metabolism enhances gold nanoparticle radiosensitisation in HPV-negative head and neck cancer
Creators: name: Feng, Jie
name: Pathak, Varun
name: Byrne, Niall M.
name: Wang, Tongchuan
name: Yin, Cancan
name: Medina, Reinhold J.
name: Coulter, Jonathan A.
Publisher: name: figshare
Year: 2024
Subjects: Biochemistry
Medicine
Molecular Biology
Pharmacology
Cancer

description: Additional file 1: Table S1. Sensitiser enhancement ratio (SER) and dose enhancement factor (DEF) of HNSCC cells after treatment with AuNPs, atovaquone alone or the combination under normoxia (21% O2). SER represents area under curve versus radiation only whilst DEF value demonstrates radiation enhancement at a single dose point versus radiation only. α and β component of linear quadratic has also been presented. Table S2. Sensitiser enhancement ratio (SER) and dose enhancement factor (DEF) of HNSCC cell after treatment with AuNPs, atovaquone alone or the combination under hypoxia (0.5% O2). SER represents area under curve versus radiation only whilst DEF value demonstrates radiation enhancement at a single dose point versus radiation only. α and β component of linear quadratic has also been presented. Figure S1. Intracellular uptake and localisation of AuNPs within FaDu, CAL27 and CAL33 cells under hypoxia (0.5% O2) using Cytoviva darkfield/hyperspectral imaging. Cells were treated with AuNPs for 24 h, fixed and mounted with DAPI before imaging. Representative images include enhanced dark field and the corresponding hyperspectral images, overlaid with a spectral angle map (SAM) representing AuNPs in red. Figure S2. Bioenergetic profile of HNSCC cells treated with atovaquone under hypoxic (0.5% O2) conditions. Changes in oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were monitored at 37 °C for 3 h. The resulting effect of atovaquone on OCR and ECAR were shown as a percentage of the baseline measurement after each treatment. Data shown are the means ± SEM (n=5 per treatment group). Figure S3. SF0 value of FaDu, CAL27 and CAL33 cells after treatment with AuNPs (100 μg) and/or atovaquone (AQ 30 μM) alone or in combination under both normoxia (21% O2) and hypoxia (0.5% O2). Figure S4. Flow cytometry analysis for cell cycle distribution of FaDu, CAL27 and CAL33 cells using propidium staining after treatment with AuNPs (100 μg) and/or atovaquone (AQ 30 μM) alone or in combination with radiation (4 Gy) under both normoxic (a) and hypoxic conditions (b). Experiments were performed in triplicate. Figure S5. Representative Annexin-V/-FITC/PI flow cytometry analysis of FaDu, CAL27 and CAL33 cells after treatment with AuNPs (100 μg) and/or atovaquone (AQ 30 μM) alone or in combination with radiation (4 Gy) under both normoxic (a) and hypoxic conditions (b). Experiments were performed in triplicate. Figure S6. A Representative immunofluorescence images of 53BP1 foci in CAL33 cells 24 h post radiation (2 Gy) under normoxia (21% O2) and hypoxia (0.5% O2). Cells were treated with either with AuNPs (100 μg), atovaquone (AQ, 30 μM) or the combination. Panels b and c represent mean 53BP1 foci 24 h post radiation (0 or 2 Gy) in CAL33 cells after treatment with AuNPs, atovaquone or the combination under normoxia (b) and hypoxia (c). Data presented are from at least three independent replicates performed in triplicates ± SEM. One way ANOVA followed by Turkey’s multiple comparison test was used to compared within different treatment. * represents significant difference at p&lt;0.05, ** at p&lt;0.01, *** at p&lt;0.001. Figure S7. A Representative immunofluorescence images of 53BP1 foci in CAL27 cells 24 h post radiation (2 Gy) under normoxia (21% O2) and hypoxia (0.5% O2). Cells were treated with either with AuNPs (100 μg), atovaquone (AQ, 30 μM) or the combination. Panel b and c represent mean 53BP1 foci 24 h post radiation (0 or 2 Gy) in CAL27 cells after treatment with AuNPs, atovaquone or the combination under normoxia (b) and hypoxia (c). Data presented are from at least three independent replicates performed in triplicates ± SEM. One way ANOVA followed by Turkey’s multiple comparison test was used to compared within different treatment. *Represents significant difference at p&lt;0.05. Figure S8. A Representative immunofluorescence images of 53BP1 foci in FaDu, CAL27 and CAL33 cells 2 h post radiation (2 Gy) under hypoxia (0.5% O2). Cells were treated with either with AuNPs (100 μg), atovaquone (AQ, 30 μM) or the combination. Panel b, c and d represent mean 53BP1 foci 24 h post radiation (0 or 2 Gy) in FaDu (b), CAL27 (c) and CAL33 (d) cells after treatment with AuNPs, atovaquone or the combination under hypoxia. Data presented are from at least three independent replicates performed in triplicates ± SEM. One way ANOVA followed by Turkey’s multiple comparison test was used to compared within different treatment. **Represents significant difference at p&lt;0.01.