Chapter 2 │ Page 52 transduced SC263T cells were significantly higher compared to the parental cells for both low (0.15 ± 0.03 fold; p = 0.0079) and high passages (0.21 ± 0.03 fold; p = 0.0001). Furthermore, following continuous culturing, the transduced SC263T-PH cells displayed a significant further elevation in basal ROS levels compared to their younger counterpart, SC263T-PL (Appx 2.B). Taken together, it appears that baseline intracellular ROS levels increased following fluorescence transduction, which could partially contribute to redox dysregulation and NTP sensitization. Moreover, in the context of NTP treatment, while cell survival was not a ected by fluorescence transduction at low cell passages (Figure 3a), these data highlight that there are already early changes to cellular physiology (Figure 5b-d), which must be carefully considered in the experimental design. Figure 5: Cancer cell proliferation and basal ROS level analysis. (a) Phase contrast images taken at 10x show that non-transduced cell lines, regardless of low or high passage, were more confluent at 72 h compared to their transduced counterparts. Scale bar indicates 300 µm. (b) Quantification of cell confluence revealed that the growth rates of the
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