Growth rate of single isotype strains and TUB1 mRNA transcript levels measured by the predesigned TaqMan or custom assays. (a) Spotting growth assay. Log-phase cultures of the indicated cell type were serially diluted (fivefold), spotted onto rich media, and incubated at the indicated temperature for 3 d. (b–d) Growth rate of the indicated cell types in liquid media at 24°C (b), 30°C (c), and 37°C (d). Graphs represent mean ± SEM from two trials. (e)TUB1 mRNA levels in WT (black) and Tub1-only (gray) cells using either the predesigned TaqMan assay (Thermo Fisher Scientific) or a custom probe/primers assay. The experiment was repeated four times with the TaqMan assay and twice with the custom assay. Each repetition was performed with an independent culture and RNA/cDNA preparation. Results are shown as mean ± SEM; n.s., not significant by unpaired, two-tailed Student’s t test.