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. 2019 Feb 13;38(6):e99363. doi: 10.15252/embj.201899363

Figure EV4. Analysis of the expression of RsaG and RsaD in response to stress.

Figure EV4

  1. Genomic organization of the uhpT locus in HG001 strain. This locus is conserved in all S. aureus strains (Geissmann et al, 2009). The transcription of uhpT containing RsaG is induced by the two‐component system HptRS, which senses the extracellular concentration of glucose‐6 phosphate (G‐6P).
  2. Northern blot experiment showing RsaG (left panel) or RsaI (right panel) expression in the HG001 strain. Total RNA was extracted at 2, 4, and 6 h of growth in BHI or MHB with or without addition of glucose‐6 phosphate (+G‐6P). 5S rRNA was used as loading control using the same samples, which have been migrated on another agarose gel.
  3. Northern blot experiment showing RsaG expression in the HG001∆hptRS deleted mutant. Total RNA was extracted at 2, 4, and 6 h of growth in BHI with or without addition of glucose‐6 phosphate (+G‐6P). 5S rRNA was used as loading control using the same samples, which have been migrated on another agarose gel.
  4. The β‐galactosidase activity (Miller Units) have been measured from PrpoB::fn3K::lacZ, PrpoB::fn3K::lacZ::rsaI, and PrpoB::HG001_fn3K::lacZ::rsaI mut4 expressed in HG001 and HG001ΔrsaG strains. The β‐galactosidase activity was normalized for bacterial density and the results represented the mean of four independent experiments. ****P < 0.0001, ns: not significant.
  5. Northern blot experiment showing the steady‐state level of RsaD. On the left, RsaD expression was analyzed at 2, 4, and 6 h of growth, in HG001 wild‐type strain and in ∆saeRS and ∆srrAB mutant strains. On the right, the expression of RsaD was monitored in response to nitric oxide (NO) stress. The culture performed in BHI medium until OD600 = 0.2 at 37°C was then treated with the addition of 100 μM Na‐diethylamine (+DEA NONOate). 5S rRNA was used as loading control using the same samples, which have been migrated on another agarose gel.

Source data are available online for this figure.