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. 2022 Nov 23;13:1026780. doi: 10.3389/fmicb.2022.1026780

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Copyright © 2022 Choi, Heo and Chung.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Flr1 is required but not sufficient for the increase of Yap1-dependent caffeine tolerance. (A) 10-fold serial dilutions of yeast mutant strains were spotted on YEPD media containing caffeine and incubated for 3 days at 30°C. (B) RT-PCR analysis of YAP1 and FLR1 expressed in WT, yap1, flr1, and strains overexpressing YAP1 or FLR1. ACT1 as a loading control. The numbers of PCR cycles used are 28 (YAP1), 22 (FLR1) and 25 (ACT1), respectively. (C) 10-fold serial dilution of yeast strains with YAP1 or FLR1 overexpression were spotted on SD/-URA media containing diamide and incubated for 3 days at 30°C. (D) 10-fold serial dilutions of yeast mutant strains with FLR1 overexpression plasmids were spotted on SD/-URA media containing caffeine and incubated for 3 days at 30°C.