Fig. 3

The graphical representation of the viability comparison of strains carrying the V5 alleles against the control rad4-116 strain across the 24-h timecourse after the addition of thiamine to the media. For the strains carrying both the V5 alleles, deletions of the wildtype genes and the rad4-116 gene, the western blots of the depletion of V5 tag are shown. a Among all the hip1-V5 strains, only SPSC 1103 showed a gradual loss of viability dropping from 99% viability at 0 time point to 51% after 12 h, then to 39% viability at 24 h. All the other strains represented showed no significant loss of viability across the 24-h timecourse ranging from 100 to 91% viability. The western blot of SPSC 1103 shown represents the samples after being probed with anti-V5 (top signal) and anti-TAT1 (bottom signal) as control. The 0- and 4-h samples expressed the V5 tag while there was no visible signal for the 8-, 12- or 24-h samples. The TAT1 signal showed no signs of depletion across all 5 samples. b Among all the swi1-V5 strains, only SPSC 1099 showed a gradual loss of viability dropping from 96% viability at 0 time point to 61% after 12 h, then to 32% viability at 24 h. All the other strains represented showed no significant loss of viability across the 24-h timecourse ranging from 100 to 91% viability. The western blot of SPSC 1099 shown represents the samples after being probed with anti-V5 (top signal) and anti-TAT1 (bottom signal) as control. The 0, 4 and 8-h samples expressed the V5 tag while there was no visible signal for the 12- or 24-h samples. An additional positive control SPSC 1099 sample is shown on the right where no thiamine is added. c Among all the swi3-V5 strains, only SPSC 1102 showed a gradual loss of viability dropping from 93% viability at 0 time point to 56% after 12 h, then to 29% viability at 24 h. All the other strains represented showed no significant loss of viability across the 24-h timecourse ranging from 100 to 91% viability. The western blot of SPSC 1102 shown represents the samples after being probed with anti-TAT1 (top signal) and anti-V5 (bottom signal) as control. The 0- and 4-h samples expressed the V5 tag while there was no visible signal for the 8-, 12- or 24-h samples. An additional sample at 24 h was taken to underline the complete depletion of the V5 tag. d Among all the mrc1-V5 strains, only SPSC 1116 showed a gradual loss of viability dropping from 95% viability at 0 time point to 28% after 12 h, then to 4% viability at 24 h. All the other strains represented showed no significant loss of viability across the 24-h timecourse ranging from 100 to 91% viability. The western blot of SPSC 1116 shown represents the samples after being probed with anti-V5 (top signal) and anti-TAT1 (bottom signal) as control. The 0-h sample strongly expressed the V5 tag while the 4- and 8-h samples expressed very weak V5 signals, but there were no visible signals for the 12- or 24-h samples