Prolong gold antifade mountant plus dapi

MCF-7, HCT116, and HeLa cell lines were seeded in 8-well Nunc™ Lab-Tek™ Chamber Slide System (Thermo Fisher Scientific) at 35 × 10⁴ cells/well together with 0.25, 0.5, and 1 mg/mL of each β-Nb₂ZnO₆ (A) and (B) nanoparticles for 48 h. The cells were fixed using cold absolute methanol for 10 min, then treated with ProLong™ Gold Antifade Mountant plus DAPI (4′,6-diamidino-2-phenylindole) from TFS. The illustrations were recorded with an LSM 700 confocal microscope (Zeiss, Jena, Germany).

UM cell lines were fixed in PBS/4% paraformaldehyde for 10 min, washed 3 times for 5 min each in PBS, blocked and permeabilised in PBS/1% BSA/10% Normal Donkey Serum (NDS)/0.025% TX-100 for 20 min, then incubated overnight at 4 °C with primary antibody (Table S1) diluted in PBS/1% BSA/1% NDS and washed 5 times for 5 min each in PBS/1% BSA/0.025% TX-100. Cells were then incubated for 1 h at RT with Alexa Fluor 488 conjugated donkey anti-rabbit or anti-mouse IgG (Abcam, Table S1) diluted 1:500 in PBS/1% BSA/1% NDS, washed 3 times for 5 min in PBS/0.025% TX-100, mounted with ProLong Gold Antifade Mountant plus DAPI (ThermoFisher Scientific, Waltham, MA, USA) and analysed using an Olympus BX51 microscope equipped with an Olympus DP71 camera and DP Manager Software (Olympus, Shinjuku, Japan).