A. Schematic diagram describing the procedures of the photochemical activation of membrane lipid peroxidation (PALPv1) technique. BODIPY-C11 at a concentration of 5 μM was added to the cells approximately 30 min prior to imaging. Then a region in the cell was targeted by 5 pulses of the 405 nm laser. Each pulse lasts for 1 second. The 488 nm channel intensity was then recorded for 25 seconds at 1-second intervals to monitor the oxidized BODIPY-C11 signal.
B. Representative fluorescent images showing the time course of the reduced and oxidized BODIPY-C11 signals before and after the application of 405 nm laser pulses in wildtype 786-O cells. Scale bar indicates 10 μm. Five independent experiments were performed.
C. Representative fluorescent images showing the reduced and oxidized BODIPY-C11 signal before and after the application of 405 nm laser pulses in primary cortical neurons after 2-day culture in vitro. Scale bar indicates 10 μm. This experiment was performed once and 9 neurons were analyzed.