Hanne Verswyvel

Chapter 2 │ Page 50 Figure 4: Light excitation from di erent sources results in altered NTP responses and cellular physiology. (a) After exposure to ambient light, no significant di erences in NTP sensitivity were reported for parental cells (SC263), compared to cells incubated in the dark. (b) In contrast, transduced cells (SC263T) became more sensitive to high NTP treatment energy following prolonged ambient light exposure. (c) Repetitive, time-lapse imaging significantly elevated intracellular ROS levels in SC263T-PL cells compared to parental SC263-PL cells, (d) which became more pronounced over increased passaging. Data are presented as mean±SEM of 3 independent repeats (n=15). Statistical significance was determined with a Two-Way ANOVA and a post hoc Tukey’s multiple comparison test for cell survival analysis; a mixed-e ect model and the Holm-Sidak’s multiple comparison test was used for the assessment of intracellular ROS levels. * p ≤ 0.05, *** p ≤ 0.001. 3.3 Fluorescently-Transduced Cells Acquire a Lower Rate of Proliferation and Higher Baseline Intracellular ROS Levels It has been hypothesized that NTP therapy can preferentially target fastproliferating cells, and therefore, we tested whether the increased NTP sensitivity of the SC263T-PH cell line was due to higher growth rates compared to the other cell counterparts. The cells were seeded into 96-well plates and monitored for 72 hours with phase contrast live-cell imaging. Interestingly, the non-transduced cell lines, regardless of low or high passages, proliferated at a faster rate compared to the transduced cell lines (Figure 5a). Quantification of cell confluence revealed that following

RkJQdWJsaXNoZXIy MTk4NDMw