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Zhou, Q. and Li, Z. (2014). Polarized assembly and assembly-associated Stattic DNA Synthesis inhibitor Paclitaxel protein stability on the
falgellum attachment zone filament in Trypanosoma brucei. The 25th
Molecular Parasitology Meeting,
Conference proceedings, Woods Hole, MA, pp268.
Zhou, Q., Liu, B., Sun, Y. and He, C. Y. (2011). A coiled-coil- and C2-domain-containing protein is
required for FAZ assembly and cell morphology in Trypanosoma brucei. J Cell Sci 124, 3848-58. Fig. 1. FAZ2 is a new FAZ protein needed for flagellum adhesion. (A) FAZ2 localizes to your FAZ
filament. Cells expressing FAZ2::3HA were co-immunostained with FITC-conjugated anti-HA mAb and
anti-FAZ1 mAb or anti-CC2D pAb. Scale bars: 5 ��m. (B) Depletion of FAZ2 by RNAi.
endogenously tagged with a triple HA epitope in cells harboring the FAZ2 RNAi construct. Cells had been handled with PEME plus 1% NP40, and cytosolic (S) and cytoskeletal (P) fractions had been separated and
detected by immunoblotting Stattic DNA Synthesis inhibitor Paclitaxel with anti-HA antibody. Exactly the same blot was re-probed with ��-tubulin and
TbPSA6 as cytoskeleton and cytosol markers, respectively. (C) FAZ2 knockdown inhibits cell
proliferation. (D) Quantification of cells with distinctive numbers of nucleus (N) and kinetoplast (K) upon
FAZ2 RNAi. Error bars represent S.D. calculated from 3 independent experiments. (E) Percentage of
cells with detached flagella upon FAZ2 RNAi. Error bars represent S.D. from 3 independent
experiments. Fig. 2. FAZ2 depletion destabilizes FAZ proteins. (A, D). Immunostaining of FAZ1 and
3330::3HA in manage and FAZ2 RNAi cells. Tb927.73330 was tagged at its endogenous locus
using a C-terminal triple HA epitope in FAZ2 RNAi cells. White arrows indicate the quick, new FAZ
filament (snFAZ) connected using the detached new flagellum (black arrows). Scale bars: 5 ��m. (B, E).
Ranges of FAZ1 (B) and Tb927.7.3330 (E) in FAZ2 RNAi cells. The two genes were each and every endogenously
tagged that has a triple HA epitope in FAZ2 RNAi cells, and detected by anti-HA mAb. The exact same blot was
probed with ��-tubulin and TbPSA6 as cytoskeleton and cytosol markers, respectively. Protein band
intensity was established, normalized with that of loading controls, and plotted as histograms proven
beneath the western blots. Error bars signify S.D. calculated from three independent experiments.
cytosolic fraction; P: cytoskeleton fraction. (C, F). Stability of 3HA-tagged FAZ1 and Tb927.7.3330 in
FAZ2 RNAi cells. FAZ2 RNAi was Stattic DNA Synthesis inhibitor Paclitaxel induced with tetracycline for 72 h or with tetracycline for 72 h and MG-132 for 8 h. FAZ1 and Tb927.7.3330 have been detected by anti-HA mAb. Exactly the same blot was re-probed
with anti-��-tubulin and anti-TbPSA6, respectively. Protein band intensity was established as described
above. Error bars indicate S.D. calculated from 3 independent experiments. S: cytosolic fraction; P:
cytoskeleton fraction. Fig. 3.