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. 2020 Oct 9;10:574861. doi: 10.3389/fonc.2020.574861

Figure 4.

Figure 4

The effect of sapitinib on the cellular location of ABCB1 in SW620/Ad300 cancer cells. ABCB1-overexpressing SW620/Ad300 cells were seeded in 24-well plate and incubated with or without sapitinib (5 µM) for 0, 24, 48 and 72 h. The cells were then fixed using 4% paraformaldehyde at 37°C for 5 min. Subsequently, the cells were blocked using 6% BSA at 37°C for 1 h and co-cultured with the monoclonal antibody C219 at 4°C overnight. The secondary antibody, Alexa Fluor 488 was added to co-culture with the cells and incubated at 37°C for 1 h. After washing with ice-cold PBS, the nuclei were counterstained with DAPI at 37°C for 10 min. An EVOS® FL Auto fluorescence microscope was used to detect and capture the immunofluorescence images. SW620 cells were used as a negative control.