Figure 5

Thd14 expression peaks in pre-meiotic stages. a Synchrony of conjugating cells used for cDNA analyses. Cells of two different mating types were mixed to initiate conjugation, and samples of the conjugating culture were dried and stained with DAPI for visualization of stages by fluorescence microscopy every hour. Hours post mixing are indicated ("hrs") above the corresponding stages illustrated. The percentage of conjugating pairs in each stage at the corresponding time is indicated below the diagram. The last stage is percentage of single cells. b Relative THD14 expression of starved CU428 (CU Stv) and SB1969 (SB Stv) as well as during conjugation (0-14 and 24 hours after mixing) was determined using quantitative real-time PCR. THD14 expression was normalized to CYP1 expression (THD14/CYP1) and the levels of THD14 expression were set relative to the starved CU428 (normalized values of each sample/CU428). Error bars represent the standard deviation of three separate trials and asterisks indicate time points that had a significant (p < 0.05) increase in expression of THD14 compared to either the starved CU428 cells (CU Stv) or immediately after mixing (0 hrs). THD14 mRNA levels increased early in conjugation at (1-3 hours) and late (24 hours) after completion of conjugation. c Relative THD14 expression of logarithmically growing CU428 ("Log") as well as during starvation (hours 0 through 4) was determined using quantitative real-time PCR. THD14 expression was normalized to that of BTU1 and the levels of THD14 expression were set relative to logarithmically growing cells. Error bars represent the standard deviation of three separate trials and asterisks indicate time points that had a significant (* p < 0.05, *** p < 0.005) increase in expression of THD14 compared to logarithmically growing cells.