Brassinosteroids (BRs) are steroid hormones that play an essential role in plant growth and development. However, the contradictory results of previous studies make their role in cell division unclear. Using a cDNA array, we identified genes that respond to BR in the det2 suspension culture of Arabidopsis, and found that epi-brassinolide upregulated transcription of the CycD3, a D-type plant cyclin gene through which cytokinin activates cell division. RNA gel-blot analysis and cell culturing showed that epi-brassinolide may promote cell division through CycD3, and can substitute cytokinin in culturing of Arabidopsis callus and suspension cells. The CycD3 induction by epi-brassinolide was further shown to involve de novo protein synthesis, but no protein phosphorylation or dephosphorylation. Induction was also found to occur in cells of a BR-insensitive mutant, bri1, suggesting that BR induces CycD3 transcription through a previously unknown signal pathway in plants.