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. 2016 Aug 11;113(35):9870–9875. doi: 10.1073/pnas.1602455113

Fig. S4.

Fig. S4.

CsrAI14M mutant is resistant to inhibition by FliW but still binds FliW in vivo. (A) Genetic neighborhood containing the fliW, csrA, and hag genes. Large open arrows represent ORFs (lengths not to scale). Bent arrows indicate promoters. Horizontal lines indicate the boundaries of the in-frame deletion mutations corresponding to the lane numbers of Western blots shown in B and C. (B and C) Western blots of whole-cell lysates grown in the presence of 1 mM IPTG and crosslinked with 0.3% formaldehyde were developed using primary antibodies against CsrA (B) and FliW (C), respectively. All lanes contain a deletion of the native csrA gene. Some lanes are also deleted for either fliW and/or hag, as indicated. CsrA proteins indicated above the line were expressed in trans or no CsrA protein was provided as indicated by the null symbol (Ø). The following strains were used to generate lysates for both panels: lane 1, ΔfliWcsrAhag (DK1483); lane 2, ΔcsrA Physpank-csrA (DK1522); lane 3, ΔcsrAfliW Physpank-csrA (DK1469); lane 4, ΔcsrAhag Physpank-csrA (DK1503); lane 5, ΔfliWcsrAhag Physpank-csrA (DK3608); lane 6, ΔcsrA Physpank-csrAN55D (DK3005); lane 7, ΔfliWcsrA Physpank-csrAN55D (DK3011); lane 8, ΔcsrAhag Physpank-csrAN55D (DK3628); lane 9, ΔfliWcsrAhag Physpank-csrAN55D (DK3614); lane 10, ΔcsrA Physpank-csrAI14M (DK3004); lane 11, ΔfliWcsrA Physpank-csrAI14M (DK3010); lane 12, ΔcsrAhag Physpank-csrAI14M (DK3627); and lane 13, ΔfliWcsrAhag Physpank-csrAI14M (DK3613).