Crl effect on RpoS levels depends on RpoS and RssB.

A) The fluorescence of the degradable RpoS-mCherry fusion in WT (NM801), ΔrpoS (SB238), rssA2::cm (SB226), Δcrl (SB228), ΔrpoS rssA2::cm (SB281), ΔrpoS Δcrl rssA2::cm (SB280) and Δcrl rssA2::cm (SB282) strains was measured as described in Fig 4A. B) The fluorescence of the degradable RpoS-mCherry fusion in WT (NM801), ΔrpoS (SB238), ΔrpoS Δcrl (SB239), Δcrl (SB228), ΔrssB Δcrl (SB283), and ΔrssB (SB225) strains was measured as described in Fig 4A. C) Ratios between RFU values in strains crl^-/ crl⁺ in WT, rpoS^- and rssB^- strains, from the data shown in S7B Fig. WT is set to 100. D) Full graphs of the ratios between the strains deleted of crl (Δcrl (SB228), Δcrl ΔrpoS rssA2::cm (SB280) and Δcrl rssA2::cm (SB282)) and the corresponding crl⁺ strains (WT (NM801), ΔrpoS rssA2::cm (SB281) and rssA2::cm (SB226)). Fluorescence and data analysis were performed as described in Fig 4A, with data during growth from S7A Fig. WT is set to 100. E) The fluorescence of the non-degradable RpoS-mCherry fusion in WT (NM802), ΔrpoS (SB439) and Δcrl (SB437) strains was measured as described in Fig 4A. F) The fluorescence of the chromosomally-inserted PgadB-mCherry transcriptional fusion from WT (SB66), ΔrpoS (SB67), ΔrssB (SB242), Δcrl (SB243), ΔrpoS ΔrssB (SB298), ΔrssB Δcrl (SB297) and Δcrl ΔrpoS (SB296) strains was measured as described in Fig 4A. G) The effect of the absence of Crl on PgadB-mCherry. mCherry fluorescence of the strains WT (SB66), ΔrssB (SB242), Δcrl (SB243) and ΔrssB Δcrl (SB297), all containing the chromosomally-inserted PgadB-mCherry fusion was measured over time as described in Fig 4A. Using the data from rpoS⁺ strains in S7F Fig, the RFU ratios during growth between the strains containing deletion of crl compared to crl⁺, in WT or ΔrssB strains was calculated, with the value in the crl⁺ strain across growth set to 100. The arrows indicate the ratios at times 200 and 800 minutes.

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