Visualizing Mitochondrial Dynamics in Cancer Cells

CRC cells were plated in confocal dishes and then incubated with Mito-tracker (100 nM) (Thermofisher, Waltham, MA, USA) for 30 min at 37 °C. The cells were fixed with 4% paraformaldehyde for 15 min as previously described,^{34 (link)} then washed twice with PBS and permeabilized with 0.2% Triton X-100 in PBS for 15 min at room temperature. Blocking with 5% bovine serum albumin at room temperature for 45 min and incubation with primary antibody at 4 °C overnight. Follow washing for 2 times, the dishes were incubated with Alexa Flour 647 or 488‐labeled secondary antibody, respectively for 1 h. DAPI staining (1 ug/mL,10 min) was performed after two washes and observed on a Zeiss laser scanning confocal microscope (LSM880, Germany) or an ultra-high resolution microscope (Nikon N-SIM).