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. 2016 Oct 19;12(10):e1005949. doi: 10.1371/journal.ppat.1005949

Fig 6. Methylfolate trap-mediated thymineless death.

Fig 6

(A) Cellular levels of methionine, SAM, and SAH in S. typhimurium cells following SMZ treatment. metH(-) (blue triangle) displayed a lower level of methionine but higher levels of SAM and SAH than its parent metH(+) (red circle). (B) Higher level of glycine in metH(-) (blue triangle) compared to metH(+) (red circle). (C) Dynamics of nucleotide pool in S. typhimurium cells during SMZ treatment. While the levels of nucleotides and intermediates were sharply reduced in response to SMZ in metH(+) (red circle), cells of metH(-) (blue triangle) failed to deplete these metabolites. (D) Thymine abolishes SULFA-induced cell death and restores growth in metH(-). Salmonella cultures were 10X serially diluted and 5 μl of diluted cultures were spotted on LB agar in the absence or presence of 125 μg/ml SMZ and 2 mM thymine. Growth on test plates (top panel) was recorded after 24 h of incubation at 37°C. Corresponding 24-hour viability of colonies grown from spotted OD0.001 cell suspensions (arrow) was determined by measuring c.f.u. and normalized to c.f.u. values of the input inoculum (lower panel). The y-axis represents c.f.u. fold-change on a log10 scale. Bars represent standard deviations from biological triplicates. *** p<0.001; **** p<0.0001.