Cellular microtubules and DNA were visualized in DU 145 cells by indirect immunofluorescence, as previously described.14 (link) The cells were treated with 5 times the IC50 of 1, 2, 3 , or 6 or 2 times the IC50 concentration of 4 for 16–18 h; then the cells were fixed with ice-cold methanol, and microtubules were visualized with a β-tubulin antibody (T4026, Sigma- Aldrich) and a FITC-conjugated secondary antibody (F3008, Sigma-Aldrich). The DNA was visualized with 4',6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich). Images were obtained using a Nikon Eclipse 80i microscope and NIS Elements Advanced Research imaging software (Nikon Instruments, Melville, NY, USA).
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Visualizing Microtubules and DNA in DU 145 Cells
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Other organizations : University of Oklahoma, The University of Texas Health Science Center at San Antonio
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Variable analysis
independent variables
- 5 times the IC50 of compound 1
- 5 times the IC50 of compound 2
- 5 times the IC50 of compound 3
- 5 times the IC50 of compound 6
- 2 times the IC50 concentration of compound 4
- Cellular microtubules visualization
- DNA visualization
- DU 145 cells
- Indirect immunofluorescence method
- 16-18 hours of treatment duration
- Methanol fixation
- β-tubulin antibody (T4026, Sigma-Aldrich) for microtubule visualization
- FITC-conjugated secondary antibody (F3008, Sigma-Aldrich) for microtubule visualization
- DAPI (Sigma-Aldrich) for DNA visualization
- Nikon Eclipse 80i microscope and NIS Elements Advanced Research imaging software (Nikon Instruments, Melville, NY, USA) for imaging
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