Oil immersed 60 objective

Fluorescence microscopy and live imaging were performed as described previously^{15 (link)}. Briefly, an FV1200-IX83 laser scanning confocal microscope with an oil-immersed 60 × objective (Olympus, Japan) was used for fluorescence microscopy. For live cell imaging, cells grown on a glass-bottomed dish were placed on a stage top incubator (Tokai Hit, Japan) that maintained a humidified atmosphere of 5% CO₂ at 37 °C. Images were captured and analyzed using FLUOVIEW software (Olympus). Fluorescence intensity of a region of interest (ROI) was quantified using FLUOVIEW software.

Fluorescence microscopy was carried out as described previously^{35 (link)}. In brief, an FV1200-IX83 laser scanning confocal microscope with an oil-immersed 60× objective (Olympus, Japan) was used for fluorescence microscopy. For live cell imaging, cells grown on a glass-bottomed dish were placed on a stage top incubator (Tokai Hit, Japan) that maintained a humidified atmosphere and 5% CO₂ at 37 °C. Images were captured and analyzed using FLUOVIEW software (Version 4.1, Olympus).

Cells in a glass-bottomed dish were stained with 1 µg/mL Hoechst 33342 (H342, Dojindo Laboratories, Kumamoto, Japan) and 0.25 µM MitoRed (R237, Dojindo Laboratories). A glass-bottomed dish was placed on a stage-top incubator (Tokai Hit, Fujinomiya, Japan) that maintained a humidified atmosphere and 5% CO₂ at 37 °C. Fluorescence microscopy was carried out by using an FV1200-IX83 laser scanning confocal microscope with an oil-immersed 60× objective (Olympus, Tokyo, Japan). Hoechst 33342 was excited with 405 nm laser, and fluorescence at 460 nm was monitored. MitoRed was excited with 559 nm laser, and fluorescence at 580 nm was observed. Images were captured and analyzed using FLUOVIEW software (Version 4.1, Olympus, Tokyo, Japan).