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. 2011 Jan 28;77(6):2141–2152. doi: 10.1128/AEM.02347-10

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FIG. 6. — In vitro-synthesized AI-2 increases soluble GroEL level. (A) The scheme depicts the synthesis and fractionation of AI-2 and its addition to cell cultures of MDAI2 (no plasmid). First, AI-2 was synthesized in vitro from substrate SAH. Second, any unreacted SAH and by-products homocysteine and adenine were removed by HPLC. After mobile-phase solvent removal via vacuum pump, the fractionated AI-2 was added to MDAI2 cell cultures. (B to D) Samples were taken at 1 h.p.i. Chaperones GroEL (soluble [B] and insoluble [C] fractions) and DnaK (soluble fraction [D]) were analyzed by Western blotting. The results shown here are representative of duplicate experiments and triplicate assays. (E and F) Transcriptional analysis of groEL (E) and dnaK (F) indicating no change due to the addition of AI-2. Error bars show standard error. Western blotting and RT-PCR were performed within the linear ranges of the assays.