Polyphosphate Maintains Intestinal Homeostasis Figure 1. HSP27-inducible substance was separated from L. brevis SBC8803 culture supernatant. (A) HSP27 induction from Caco2/BBE cells by culture supernatant obtained from the L. brevis 8803 culture broth at 12, 36 and 60 h. (B) HSP27 induction from Caco2/BBE cells by culture supernatant of L. brevis 8803, L. brevis 8013 and LGG obtained from the bacterial culture broth at 6 and 24 h. (C) HSP27 induction by filtrate of L. brevis SBC8803 culture supernatant passed through a 5-, 10-, 30- or 50-kDa membrane. (D) HSP27 induction by 65% or 90% ammonium sulfate precipitate of L. brevis SBC8803 culture supernatant. (E) HSP27 induction by fraction separated from 65% ammonium sulfate precipitate by DEAE anion-exchange chromatography. (F) HSP27 induction by fraction separated from 0.1 M NaCl-eluted fraction by size-exclusion chromatography. (G) HPLC chromatogram of the HSP-inducible fraction. Sample was separated on a Shodex KW800 column (300 mm68 mm ID), eluted with 20 mM phosphate buffer (pH 6.5) at a flow rate of 0.1 mL/min. The eluent was monitored with an ultraviolet spectrophotometer at 220 nm. (H) HSP27 induction by 10 mg/mL of HSP27-inducible fraction (n = 3). doi:10.1371/journal.pone.0023278.g001 PLoS ONE | www.plosone.org 3 August 2011 | Volume 6 | Issue 8 | e23278