Fig. 4.

Generation of recombinant APH_1235 and antisera against APH_1235. (A and B) Whole-cell lysates of uninduced E. coli (lane U), E. coli induced (lane I) to express GST-1235 (A) or His-1235 (B), and GST-1235 (44.2 kDa) purified (lane P) by glutathione-Sepharose affinity chromatography (A) or His-1235 (13.5 kDa) purified by Ni²⁺ affinity chromatography (B) were separated by SDS-PAGE and stained with Coomassie blue. (B) Western blot (WB) analysis was performed using mouse anti-APH_1235 (raised against GST-1235) to detect His-1235 (lane WB). Black arrows denote GST-1235 (A) and His-1235 (B). The white arrow in panel B denotes what is presumably a His-1235 dimer. (C) Western blot analysis in which anti-APH_1235 was used to screen whole-cell lysates derived from uninfected HL-60 cells and A. phagocytophilum organisms (lane Ap).