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. 2018 Jan 4;46(3):1345–1361. doi: 10.1093/nar/gkx1284

Figure 6.

Figure 6.

CspA expression and auto-regulation require the cspA 5′UTR. (A) cspA3xFLAG mRNAs generated from pCspA3xFLAG and pCspA3xFLAG-Δ5′UTR plasmids in the WT and ΔcspA strains. Total RNA extraction was performed at mid-exponential phase after growth at 37°C and 200 rpm. Samples were run into 1.25% agarose and transferred to Nitran membranes. Northern blots were then developed using a 32P-labeled anti-FLAG oligo probe. Ethidium bromide staining of 16S RNA is shown as a loading control (LC). Processed cspA3xFLAG mRNA is indicated with an asterisk. (B) CspA3xFLAG protein levels from the pCspA3xFLAG and pCspA3xFLAG-Δ5′UTR plasmids in the WT and ΔcspA strains. Total protein extraction was performed at mid-exponential phase, after growth at 37°C and 200 rpm. Samples were run in 12% polyacrylamide gels and transferred to Nitrocellulose membranes. The Western blot was developed using peroxidase conjugated anti-FLAG antibodies and a bioluminescence kit. Exp 1 and Exp 2 indicate two different exposition times. A Coomassie stained gel portion is shown as a loading control (LC).