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. 2020 Aug 31;16(8):e1008131. doi: 10.1371/journal.ppat.1008131

Fig 4. Conditional degradation of AID-tagged proteins is efficient in pre-erythrocytic stages.

Fig 4

A) Western blot analysis of protein lysates from CDPK1-aid-HA and CDPK5-aid-HA parasites treated with IAA or vehicle. CDPK1-aid-HA and CDPK5-aid-HA were detected using an anti-HA antibody. GFP was used as a loading control. CDPK1-aid-HA and CDPK5-aid-HA have predicted molecular weights of approximately 90 kDa. The arrowhead indicates a byproduct of partial degradation of CDPK5-aid-HA. B) Depletion of CDPK5-aid-HA protein in merosomes was examined using immunofluorescence. Optical sections of representative IAA- or vehicle-treated merosomes immunostained with an anti-HA antibody. Parasite nuclei were visualized using DAPI. C) Effects of IAA treatment on Ostir1 pre-erythrocytic stages were examined by quantifying sporozoite invasion of HepG2 cells, the number of liver stages detected at 48 h p.i. and the number of merosomes released into the media at 66–70 h p.i., in the presence or absence of IAA. Results shown are average of 2–4 experiments with 3 technical replicates (± SEM), normalized to vehicle-treated samples for each assay. Data were analyzed using an unpaired t-test, not significant (n.s.) P value > 0.05.