Author: Lin, Mingqun; Liu, Hongyan; Xiong, Qingming; Niu, Hua; Cheng, Zhihui; Yamamoto, Akitsugu; Rikihisa, Yasuko
Title: Ehrlichia secretes Etf-1 to induce autophagy and capture nutrients for its growth through RAB5 and class III phosphatidylinositol 3-kinase Document date: 2016_8_19
ID: x5y551c8_62
Snippet: HEK293 cells expressing codon-optimized Etf-1-HA and GFP-RAB5 (WT, DN, or CA) or GFP control were lysed in modified lysis buffer as described above. After centrifugation, each supernatant fraction was incubated with mouse anti-HA mAb cross-linked on protein G-sepharose beads (BioLegend, 900801) for 2 h. After extensive washing with lysis buffer, bound proteins were dissociated from the beads with 2£ SDS sample buffer, and western blotting was pe.....
Document: HEK293 cells expressing codon-optimized Etf-1-HA and GFP-RAB5 (WT, DN, or CA) or GFP control were lysed in modified lysis buffer as described above. After centrifugation, each supernatant fraction was incubated with mouse anti-HA mAb cross-linked on protein G-sepharose beads (BioLegend, 900801) for 2 h. After extensive washing with lysis buffer, bound proteins were dissociated from the beads with 2£ SDS sample buffer, and western blotting was performed.
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