Fig. 2.

Identifying genes important for filamentation for GARP complex mutants. a) To repress BCY1 expression in the tetO-BCY1/tetO-BCY1 strains, cells were grown overnight at 30°C in YPD medium in the absence or presence of 20 µg/ml doxycycline (DOX) with shaking and subsequently grown in the absence or presence of 20 µg/ml DOX for 6 h at 30°C with shaking prior to imaging. Scale bar represents 10 µm. b) Cells were grown overnight in the absence or presence of 20 µg/ml DOX. Cells were subcultured into YPD with the same DOX conditions for 5 h before pelleting, RNA extraction, cDNA synthesis, and qRT-PCR. Transcript levels of BCY1 were normalized to ACT1 and are relative to the wild-type no DOX control. Data are presented as mean ± SEM of technical triplicates. Significance was determined by unpaired t-test. c) To overexpress UME6 in the tetO-UME6/UME6 strains, cells were grown overnight at 30°C in YPD medium in the absence or presence of 20 µg/ml DOX with shaking and subsequently imaged. Scale bar represents 10 µm. d) Cells were grown as in (b). Transcript levels of UME6 were normalized to ACT1 and are relative to the wild-type no DOX control. Data are presented as mean ± SEM of technical triplicates. Significance was determined by unpaired t-test. e) Strains were grown overnight at 30°C in YPD medium with shaking and subsequently imaged. Scale bar represents 10 µm. ***P < 0.001 and ****P < 0.0001.