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By using Hoechst 33342,rabbit anti calmodulin anti-body,FITC-labeled goat anti rabbit IgG and SR101(sulforhodamine 101)simultaneously to stain individual normaland transformed cells,the microspectrophotometric analysisdemonstrated that 3 markers which represented the nucleus,calmodulin and total protein respectively,could be recog-nized in individualj cells without interference.The phase ofthe cell cycle was determined by DNA content(Hoechst33342).We found that in transformed cells(NIH3T3)tsRSV-LA90,cultured at 33℃ and transformed C3H10T1/2 cells),the ratio of calmodulin to total protein(based onthe phases of cell cycle)was higher than that in normal cells(NIH3T3 tsRSV-LA90 cells,cultured at 39℃ andC3H10T1/2 cells)in every cell cycle phase.This ratio in-creased obviously only from G_1 to S phase in either normalor transformed cells.The results showed that calmodulinreally increased during the transformation,and its increasewas specific.In the meantime when cells proceeded from G_1to S,the intracellular calmodulin content also increasedspecifically.
By using Hoechst 33342, rabbit anti calmodulin anti-body, FITC-labeled goat anti rabbit IgG and SR101 (sulforhodamine 101) to stain individual normantly transformed cells, the microspectrophotometric analysis demonstrated that 3 markers which represented the nucleus, calmodulin and total protein respectively, could be recog-nized in individualj cells without interference. The phase of the cell cycle was determined by DNA content (Hoechst 33342) .We found that in transformed cells (NIH3T3) tsRSV-LA90, cultured at 33 ° C and transformed C3H10T1 / 2 cells) the ratio of calmodulin to total protein (based on the phases of cell cycle) was higher than that in normal cells (NIH3T3 tsRSV-LA90 cells, cultured at 39 ° C and C3H10T1 / 2 cells) in every cell cycle phase. This ratio in- only from G_1 to S phase in either normal normal transformed cells. The results showed that calmodulin really increased during the transformation, and its increase was specific. The meantime when cells proceeded from G_1t o S, the intracellular calmodulin content also increasedspecifically.