Fig. 1: The levels of MIF protein fluctuate throughout cell cycle and MIF is located at basal body in RCTE cells.

a Schematic showing the cell cycle phase inhibited by nocodazole and thymidine. b Experimental outlines for using thymidine-nocodazole block or double thymidine block methods to synchronize RCTE cells. Fluorescence-activated cell sorting (FACS) was used to estimate the cell-cycle profiles in thymidine/nocodazole (c) and double thymidine block (d) treated cells by measuring the DNA contents in those cells with propidium iodide staining. Western blot analysis of whole cell lysates derived from RCTE cells synchronized in M phase by thymidine-nocodazole (e) and in late G1/S phase by double thymidine treatment (f) following by releasing back into the cell cycle. g Representative images of RCTE cells stained with MIF antibody and co-stained with γ-tubulin (γ-Tub) (red), which were visualized under 2D microscope and three-dimensional structured illumination microscopy (3D-SIM). Scale bars, 5 μm (top panels) and 500 nm (middle and bottom panels of 3D images). h The intensity plots of the rings in 3D images (g) from top and side views. i Representative images of RCTE cells stained with MIF antibody and co-stained with pericentrin (red). Scale bar, 5 μm. j Representative images of RCTE cells stained with MIF antibody and co-stained with acetylated α-tubulin (Ac-αTub) (red), which were visualized under 2D microscope and 3D-SIM microscopy. The “top or side” views are regarding the centriole structures. Scale bars, 5 μm (top panels) and 500 nm (middle and bottom panels of 3D images).