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Supplement ary Fig. S 1. a. b. IFT88#2. IFT88. GFP. siRNA:. a tub /. IFT88. IFT88. 100. 75. WB anti- IFT88. 50. WB anti- g tubulin. c. siRNA:. GFP. 14. IFT88. 10. Mitotic index (%). 6. 2. T (h). 48. 0. 3 2. d. 20. 0. -10. 10. 30. 40. 50. 60. GFP. siRNA:.

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C

Supplementary Fig. S1

a.

b.

IFT88#2

IFT88

GFP

siRNA:

a tub/

IFT88

IFT88

100

75

WB anti-IFT88

50

WB anti-g tubulin

c.

siRNA:

GFP

14

IFT88

10

Mitotic index (%)

6

2

T (h)

48

0

32

d.

20

0

-10

10

30

40

50

60

GFP

siRNA:

-10

0

40

160

220

280

300

330

IFT88

e.

60

siRNA:

GFP

IFT88

40

% cells

20

0

Time (min)

10-50

60-100

110-150

> 150


C

Phos-H3

positive cells

(Fold increase)

3

2

1

0

siRNA:

MO:

Con

IFT88

control

IFT88sc

Supplementary Fig. S2

a.

b.

90

siRNA:

IFT88sc

Control

70

72h

48h

72h

32h

48h

32h

100

50

% cells

75

30

WB anti-IFT88

75

10

Disrupted

poles

Disrupted

g tubulin

Misaligned

chrom

Spindle

tilt

WB anti-dynein

IFT88sc

siRNA:

c.

40

control

30

IFT88sc

20

% cells w/

spindle defects

GFP / a tubulin

10

0

GFP neg

GFP pos

GFP-IFT88 / LLC-PK1

d.

a tubulin

/ g tubulin

100

Tg737 +/+

80

Tg737 +/+

Tg737 -/-

60

% cells

40

20

0

Tg737 -/-

0

10

5

20

Spindle angle (a °)

a tubulin/

g tubulin/

Phos-H3

e.

Control MO

IFT88 MO


C

16

12

g tubulin intensity

(x105 a.u.)

8

4

0

siRNA:

IFT88

GFP

Supplementary Fig. S3

a.

b.

siRNA: IFT88

siRNA: GFP

EB1

EB1

14000

14000

Gray scale

12000

Gray scale

12000

10000

10000

60000

60000

5051

5051

35000

35000

Gray scale

Gray scale

10000

10000

0

10

20

0

10

20

Distance (mm)

Distance (mm)

c.

d.

IFT88/

EB1/

EB1

IFT88

DAPI

70

GFP

50

MT nucleation rate

(min-1)

siRNA:

30

10

EB1

siRNA:

GFP

IFT88


C

Supplementary Fig. S4

a tub

Dyn IC

a tub/

Dyn

a.

a tub

a tub/

Dyn

Dyn IC

a tub

IgG

a tub/

IgG

a tub/

IgG

IgG

a tub

Negative controls

a tub/

a tub/

a tub

No primary

No primary

a tub

No primary

No primary

b.

c.

Mitotic shake off

Mitotic shake off

IP: Con

Dyn IC

Input

IP: Con

IFT88

Input

100

100

75

WB anti-IFT88

WB anti-IFT88

75

75

WB anti-Dynein IC

WB anti-Dynein IC

Mitotic shake off:

Input

IP Flag

IP Ig

d.

IMCD

IMCD Flag-IFT52

+ - + - -

- + - + +

100

IFT88

WB anti-IFT88

75

Flag-IFT52

WB anti-Flag

50

e.

IP Ig

IP IFT88

Input

Mitotic shake off:

IMCD

IMCD Flag-IFT52

+ - + -

- + - +

+ -

- +

100

IFT88

75

WB anti-IFT88

Flag-IFT52

50

WB anti-Flag


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