Stattic DNA Synthesis inhibitor Paclitaxel

The
two bright green fluorescence dots on the proximal base of the flagella are KMP11 signal inside the Stattic DNA Synthesis inhibitor Paclitaxel basal
bodies. Scale bar: 5 ��m. (B) Stability of CC2D and KMP11 in FAZ2 RNAi cells. FAZ2 RNAi cell line expressing endogenously PTP-tagged KMP11 was induced with tetracycline for 72 h or with tetracycline
for 72 h and MG-132 for 8 h. Soluble and pellet fractions have been immunoblotted with anti-CC2D and anti-
Protein A to detect CC2D and KMP::PTP, respectively. The same membrane was re-blotted with anti-��-
tubulin and anti-TbPSA6. Protein band intensity was determined as described in Fig. 2, and plotted being a
histogram shown beneath the Western blots. Error bars signify S.D. calculated from 3 independent
experiments. S: cytosolic fraction; P: cytoskeleton fraction.

(C) Result of KMP11 RNAi on FAZ2 and
CC2D localization. FAZ2 was endogenously tagged using a triple HA epitope in KMP11 RNAi cell line.
Cells were co-immunostained Stattic DNA Synthesis inhibitor Paclitaxel with FITC-conjugated anti-HA mAb and anti-CC2D pAb. White
arrowheads display the FAZ2::3HA and CC2D in the proximal base from the detached new flagellum (black
arrow). Scale bar: 5 ��m. (D). Stability of FAZ2 and CC2D in KMP11 RNAi cells. KMP11 RNAi cell line expressing FAZ2::3HA was induced with tetracycline for 32 h or with tetracycline for 32 h and MG-132
for 8 h. Soluble and pellet fractions were immunoblotted with anti-CC2D pAb and anti-HA mAb to detect
CC2D and FAZ2::3HA, respectively. The identical membrane was re-blotted with anti-��-tubulin and anti-
TbPSA6 as cytoskeleton and cytosol markers, respectively.

Protein band intensity was established as
described in Fig. 2, and plotted like a histogram proven under the Western blots. Error bars indicate S.D.
calculated from three independent experiments. S: cytosolic fraction; P: cytoskeleton fraction.
Fig. 6. CC2D depletion destabilizes FAZ2 and KMP11. (A). Effect of CC2D RNAi over the localization
of FAZ2. FAZ2 was endogenously tagged using a C-terminal triple HA epitope in cells harboring a CC2D
RNAi construct. Cells had been immunostained with FITC-conjugated anti-HA mAb. The white arrow demonstrates
the FAZ2::3HA in the proximal base of detached new flagellum, along with the black arrow signifies the
detached new flagellum. Scale bar: 5 ��m. (B) Effect of CC2D RNAi on KMP11 localization. KMP11 was
endogenously tagged with an Stattic DNA Synthesis inhibitor Paclitaxel HA tag in CC2D RNAi cell line.

Cells had been co-immunostained with FITC-conjugated anti-HA mAb and anti-CC2D pAb. White arrows indicate the KMP11::HA from the FAZ
filament, whereas the white arrowhead exhibits the CC2D signal at the proximal base with the detached new
flagellum (black arrow). Scale bar: 5 ��m. (C, D). Stability of FAZ2 (C) and KMP11 (D) in CC2D RNAi
cells. CC2D RNAi cell line expressing endogenously 3HA-tagged FAZ2 or PTP-tagged KMP11 was
induced with tetracycline for 72 h or with tetracycline for 72 h and MG-132 for 8 h.