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. 2022 Jun 10;18(6):e1010089. doi: 10.1371/journal.ppat.1010089

Fig 6. Tor1 N-terminal HEAT repeats were required for oxidative stress-induced delay of translation initiation.

Fig 6

A. Cells expressing GFP from tetO (ON) (strain TETG25B) were grown in YPD medium for 15 h, then inoculated into fresh YPD and pre-grown for 3.5 h (Time 0, 0 min). GFP expression was induced with 50 μg/ml doxycycline in SC medium with Vehicle (DMSO) or 15 μM Plumbagin (Plum). Negative control (NC) cells were grown in SC medium without doxycycline (with DMSO) for 120 min. Total protein extracts were probed with antibody to GFP, and tubulin (Tub) as loading control. Dens: signal intensity ratio of GFP to Tub. Representative of 2 biological replicates. B. Cells expressing pMAL2-GFP in backgrounds with distinct TOR1 alleles were pre-grown in YPD medium with 10 ng/ml doxycycline for 15 h (+/+, JKC2616; -/+, JKC2620; Del381, JKC2624; FL, JKC2628). GFP expression was induced by inoculation into YP-Maltose medium with 5 ng/ml doxycycline, containing 13 μM Plumbagin (Plum) or DMSO as vehicle (Veh). (0 h, JKC2616 after pre-growth; N, JKC2616 inoculated into YPD instead of YP-Maltose with 5 ng/ml doxycycline, Plumbagin or DMSO, grown for 9 h). Total protein extracts were probed with antibody to GFP, and tubulin (Tub) as loading control. Dens: signal intensity ratio of GFP to Tub.