Fig. 1.

Establishment of aposymbiotic amoebae, R18DOX and S13RFP, from R18WT and S13WT, respectively, by antibiotic treatment.

(A) Representative FISH images of R18WT and S13WT amoebae before treatment with antibiotics. The amoebae were observed with a confocal laser microscope. Green, amoebal 18S rRNA; orange, endosymbiotic bacteria. 1,000× magnification. Arrows show representative bacterial clusters. (B) Representative DAPI-stained images of R18 and S13 amoebae before and after treatment with antibiotics. 1,000× magnification. The amoebae were observed with a conventional fluorescence microscope. Arrows show representative bacterial clusters. N, amoebal nucleus. (C) Representative RT-PCR products amplified from amoebae and resolved by agarose gel electrophoresis. WT, wild type; DOX, doxycycline; RFP, rifampicin.