Ehrlichia secretes Etf-1 to induce autophagy and capture nutrients for its growth through RAB5 and class III phosphatidylinositol 3-kinase

Lin, Mingqun; Liu, Hongyan; Xiong, Qingming; Niu, Hua; Cheng, Zhihui; Yamamoto, Akitsugu; Rikihisa, Yasuko

E. chaffeensis-infection in bone marrow-derived macrophages from wild-type and atg5 flox/flox -Lyz2-Cre mice BMDMs with atg5 knockout were isolated from atg5 flox/flox -Lyz2-Cre mice (atg5 TSKO), 64 in which the Cre recombinase driven by Lyz2 promoter was used to target myeloid cells to knockout the third exon of Atg5 flanked by LoxP sites. 65 Monocytes were derived from bone marrow of wild type (WT) and atg5 TSKO mice, and macrophages differentiated from the monocytes were incubated with E. chaffeensis as described. 54 The infected samples were collected at 7 d p.i and subjected to DNA isolation using the QIAamp DNA blood mini kit (Qiagen, 51106); qPCR was performed using primers specific for E. chaffeensis 16S rDNA and mouse Gapdh. 130 The protocol for using animals in this study was approved by the Institutional Laboratory Animal Care and Use Committee.