Fig. 7

Different fixation methods affect mitotic figure labeling in IMCD3 and RPE cells. IMCD3 and RPE cells were fixed with various fixation methods, and then probed with acetylated-α-tubulin to establish images of metaphase mitotic figures when subjected to different fixation methods. Separate coverslips of cells were subjected to labeling with either a membrane-bound cilia marker (AC3, Arl13b; green) or an axoneme-bound cilia marker (CSPP1, IFT20; green). All cells were stained with Hoechst for nuclear labeling (blue). A In IMCD3 cells, Arl13b did not immunolabel mitotic figures, but CSPP1, IFT20, and AC3 labeled mitotic spindles and/or centrosomes depending on the fixation method used. MeOH-treated IMCD3 cells produced the most intense centrosomal immunolabeling for CSPP1. B In RPE cells, Arl13b also did not label mitotic figures with any tested fixation method. Scale bars 10 µm