*2.8. Immunofluorescence*

The detection of CRT, HSP70, and HSP90 was performed by immunofluorescence. After the treatments, the cells were fixed with ethanol 70% (*v*/*v*, in water) at room temperature for 30 min. Next, the cells were incubated with an anti-CRT (1:75), anti-HSP70 (1:75) or anti-HSP90 (1:150) primary antibody in cold blocking buffer (2% BSA in PBS) for 1 h in an

incubator (37 ◦C), followed by washing with PBS and incubation with IgG488 secondary antibody (1:250) for 30 min at room temperature. The nuclei of the cells were labeled with DAPI. The stained cells were visualized with a fluorescence microscope (EVOS-FL, Thermo Fisher Scientific Inc., Waltham, MA, USA). The fluorescence of the stained cells was quantified with the software Photoshop CC 2015 (Adobe Systems, San Jose, CA, USA) and ImageJ 1.8.0\_172 (NIH, Madison, WI, USA).
