Fig. 2

Subcellular localization of the GroEL protein. L. interrogans serovar Copenhageni strain Fiocruz L1–130 was cultivated at 29 °C and in conditions that mimics the host environment, with shift temperatures from 29 to 37 °C for 5 h and in osmolarity of 300 mOsm. a Whole-cell lysates (W) and cell culture supernatant fractions (S) were analyzed by immunoblotting using anti-GroEL and anti-LipL31 (positive control of cytoplasmatic protein) antisera. b Whole cell (W), Triton X-114 fractions (A and D), and culture supernatant fraction (S) were analyzed by immunoblotting with anti-GroEL and anti-LipL31. c Proteinase K accessibility assay. Intact leptospires were incubated with different concentrations of proteinase K and processed for immunoblot analyses using antibodies against GroEL, LipL31 or LigA (positive control of outer membrane protein). Full-length blots are shown in the Supplementary Material as Fig. S4