Figure 4.

Effects of TDI on megalin expression. RLE-6TN cells were preincubated in serum-free medium with or without 1 µM receptor-associated protein (RAP) for 30 min and then incubated with 400 nM FITC-VDBP at 37℃ for 2 h. The fluorescence was observed using a fluorescence microscope. Scale bar, 20 µm. Fluorescence intensities of the cells were counted using an image browser, and data are presented as percent intensity (A). RLE-6TN cells were incubated with the indicated doses of TDI for 3 h, and then real-time PCR was performed (B). RLE-6TN cells were incubated with 1 mM TDI for the indicated times, and then real-time PCR was performed. The values are normalized relative to the GAPDH standard (C). RLE-6TN cells were incubated with the indicated doses of TDI for 12 h (D). RLE-6TN cells were incubated with 1 mM TDI for the indicated times, and then Western blotting analysis for megalin was performed. Tubulin was used as a loading control (E). All data are representative of three independent experiments. Values represent the means ± SEM. ^*P < 0.05, ^**P < 0.001 vs. control.