Using clear 96-well plates, cells were seeded at a concentration of 10,000 cells per well into the interior wells of the plates (columns 3-11, rows B-G inclusive) in 200 μL of serum-supplemented DMEM (MDA-MB-231 and MDA-MB-436 cells) or supplemented RPMI (A549 cells). Outer wells were filled with 200 μL of supplemented DMEM (MDA-MB-231 and MDA-MB-436 cells) or supplemented RPMI (A549 cells). Cells were incubated at 37 °C and 5% CO2 for 24 hours before photosensitiser administration. After 24 hours of incubation, supplemented media (DMEM or RPMI) was replaced with photosensitiser-containing, serum-free DMEM at various concentrations and the plates incubated for 1 hour (Verteporfin), 3 hours (5-ALA), or 24 hours (Temoporfin) under low light conditions at 37 °C and 5% CO2. The photosensitiser solution was then removed and cells were washed once with DPBS and serum-free DMEM was added to wells. Cells were then illuminated from below using LED arrays set to a power output of ~ 5 mW per LED for 15 minutes (Verteporfin) or 25 minutes (5-ALA and Temoporfin) (power output of 660 nm LEDs was limited to ~3.5 mW). 96-well plates were then incubated for 24 hours at 37 °C and 5% CO2 under low-light conditions. 24 hours post illumination, 10 μL of cell-counting kit 8 (CCK-8) substrate (Sigma-Aldrich) was added to each well and the plates incubated for 3 hours at 37 °C and 5% CO2. After 3 hours, the absorbance of each well was measured at 450 nm using a plate reader (SpectraMax M5).

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