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. 2016 Jul 18;113(31):8813–8818. doi: 10.1073/pnas.1601626113

Fig. 4.

Fig. 4.

Lpp0148 and RocR control natural competence development. (A) The L. pneumophila Paris WT strain transiently develops competence during growth at 30 °C. Expression of comEA was followed using a comEA-gfp transcriptional fusion carried by pXDC91 (GFP/OD600, green line) and natural transformability (red circle, triangle, and square) determined at different time points during growth (OD600, blue line). Error bars on natural transformation efficiencies data represent SE. (B) Natural transformability during growth at 30 °C of the L. pneumophila Paris WT, lpp0148TAA, and ΔrocR strains. (C) Expression of Lpp0148 and RocR decrease at the onset of the transformability phase at 30 °C. Expression of Lpp0148 was analyzed by Western blot, and expression of comEA and RocR was determined by Northern blot analysis. A cross-reacting band and the 5S rRNA were used as loading controls for the Western blot and Northern blot, respectively. (D) comEA mRNA half-life determination by Northern blot analysis before (OD600 of 0.9) and during (OD600 of 2.5) the competence phase. Decay of comEA was followed after transcription was stopped with rifampicin at the indicated OD. The 5S rRNA was used as a loading control.