Extended Data Fig. 10: TASH3 and LOLITA antibody development.
a) and c) The pre-immunization sera of the rabbits were tested against Col-0 lysate to ensure the absence of background immunity. The Western Blot was performed once. b) The post-immune serum, as well as the purified anti-TASH3 antibody were tested against plant lysate of a GFP-TASH3 expressing plant, as well as against purified TASH3-FLAG. The serum generates a substantial amount of background signal. The purified anti-TASH3 antibody has less background signal in the plant sample, but it still recognizes multiple bands. It works well against purified samples. The specificity in the plant lysate was confirmed via anti-GFP (magenta arrow heads). The Western Blot was performed once. d) The post-immune serum of the anti-LOLITA antibody showed a substantial amount of background signal in plant lysates, whereas the purified antibody presented only little background signal. The purified antibody was not sufficient to detect native LOLITA in Col-0 lysate, however it could detect both purified LOLITA-HIS and LOLITA-GFP expressed in planta. The Western Blot was performed once. e) The specificity of the anti-LOLITA antibody was again confirmed against anti-GFP in the LOLITA-GFP expressing sample (orange arrow heads d & e). The Western Blot was performed once.
