Ab207494

Sample preparation for western blot analysis was performed as previously described (Liu et al., 2020 (link)). Briefly, schizonts were released from erythrocytes with 0.15% saponin, resuspended with an equal volume of 2 × SDS–polyacrylamide gel electrophoresis (PAGE) protein loading buffer, and then heated for 5 min at 100°C before storing at -80°C. Proteins were separated by 10% SDS-PAGE, and then transferred to Immobilon-P transfer membranes (Millipore). Subsequent antibody incubation and membrane wash followed standard procedures. The primary antibodies used in this study included mouse anti-ty1 (Sigma, SAB4800032) at 1:1,000 and rabbit anti-aldolase (Abcam, ab207494) at 1:2,000. The horseradish peroxidase (HRP) conjugated secondary antibodies were used at 1:5,000, including goat anti-mouse IgG (Abcam, ab97040) and goat anti-rabbit IgG (Abcam, ab205718). HRP signals were detected using the ECL western blotting kit (GE healthcare). Especially, pfap2-exp2-ty1-glms parasites were tightly synchronized to a 5-h window and ring-stage parasites were diluted at 0.5% parasitemia with 2% hematocrit in the presence or absence of 5 mM glucosamine. At the second generation, 200 μL of schizont-staged samples were collected for western blotting.

The following primary antibodies were used in this study: mouse anti-Ty1 clone BB2 (Sigma-Aldrich), rat anti-HA 3F10 (Sigma-Aldrich), rabbit anti-PfMC-2TM CT and rabbit anti-PfMC-2TM SC (Bachmann et al., 2015 (link)), rat anti-RIF40 (Petter et al., 2008 (link)), rabbit anti-STEVOR PfC0025c/Pf3D7_0300400 (Bachmann et al., 2015 (link)), rabbit anti-Plasmodium aldolase (ab207494, Abcam), rabbit anti-H3 (ab1791, Abcam), rabbit anti-HP1 and rabbit anti-H2A.Z (Petter et al., 2011 (link)).