Zen 2.3 sp1 basic software

The cells were seeded on glass bottom 35‐mm dishes (MatTek Corp., Ashland, Massachusetts) containing 2 × 10⁵ cells per dish and incubated overnight. The next day, Ko143 (1 μM) was added to cells 1 hour prior to 5‐ALA addition (1.5 mM ALA, 1 hour or 4 hours) and maintained in the respective groups until imaging. The mitochondrial probe MitoTracker Green FM (100 nm) was added to the cells 15 minutes prior to live cell imaging. The cells were examined with a Zeiss LSM 880 Airyscan microscope (Carl Zeiss MicroImaging GmbH, Jena, Germany), equipped with an Ar Laser Multiline (458/488/514 nm), a DPSS 561 10 (561 nm), a Laser diode 405‐30 CW (405 nm), and a HeNe Laser (633 nm). The objective used was a Zeiss plan‐Apochromat 63xNA/1.4 oil DICII. Image acquisition, processing and analysis were performed with ZEN 2.3 SP1 basic software (Carl Zeiss), ZEN 2.3 Blue (Carl Zeiss), or Imaris 7.7.2 (Bitplane AG, Zürich, Switzerland). The Pearson's colocalization measurements were performed in ZEN 2.3 Blue with Costes' automatic thresholding.

The cells were examined with a Zeiss LSM 880 Airyscan microscope (Carl Zeiss MicroImaging GmbH, Jena, Germany), equipped with an Ar-Laser Multiline (458/488/514 nm), a DPSS-561 10 (561 nm), a Laser diode 405-30 CW (405 nm), and a HeNe-laser (633 nm). The objective used was a Zeiss plan-Apochromat 63xNA/1.4 oil DICII. The images were acquired with the Airyscan detector with a voxel size of 0.035 × 0.035 × 0.144 µm for high resolution imaging and super-resolution processing resulting in a final resolution of 120 × 120 × 350 nm. Image acquisition and processing were done with the ZEN 2.3 SP1 basic software (Carl Zeiss), or with ZEN 2.3 Blue (Carl Zeiss).