Extended Data Fig. 7: Identification of punctuate structures observed in ferroptotic HT-1080 cells stained with H4B-PMHC.
a-c. Colocalization timelapse (100x HILO) in HT-1080 cells stained with H4B-PMHC (10 nM, cyan), co-stained with LipidSpot 610 (a, magenta), lysoTracker DR (b, magenta), or mitoTracker DR (c, magenta). Imaging was conducted upon RSL3 (1 µM) addition. Scale bars = 12 µm. Scale bar of the enlarged region = 4 µm. Colocalization of all channels is shown in the “merged” panel. Each experiment was conducted once. In each experiment, 4 FOVs were monitored. d. To quantify the contribution to CTCF from these punctuate structures in Fig. 3b, the outline of these structures and that of the whole cell were identified via CellProfiler. Next, the integrated intensity and total area for punctate structures and the whole cell were determined, and intensity-time trajectories of these two regions were plotted (also see Fig. 3e). e. Regions corresponding to Golgi-associated vesicles (cyan and yellow) and lipid droplets or Golgi (magenta), were identified by CellProfiler. The vesicular structures do not colocalize with lipid droplets but were formed next to the Golgi apparatus. Scale bars = 12 µm.
