Table 2

Cell morphology and cytoskeleton organisation in PMM-depleted PC3 cells.

Group

Gene symbol

Actin

Microtubules

Cell shape

Spread area

Multi-nucleate


control

stress fibres 4%

strong patches 8%

cortical actin 12%

low 14%

processes 6%

elongated 10%

rounded 4%

larger 4%

0%

1

ARC

stress fibres 70%

NP

elongated

larger

6%

ZRANB1

stress fibres 90%

NP

elongated

larger

10%

2

C20orf142

cortical actin

NP

NP

larger 85%

NP

CAMSAP1

cortical actin, spots

NP

NP

larger 85%

NP

FMNL1

cortical actin 90%, rough edge

NP

NP

larger

NP

FAM40A

cortical actin 90%, rough edge

NP

NP

NP

NP

C21orf2

cortical actin 55%

NP

NP

NP

NP

3

FMNL2

cortical actin, spots

processes 60%

NP

larger

9%

ZNF135

spots

processes 50%

NP

NP

NP

ZMYM3

cortical actin, spots

processes 25%

NP

larger

NP

PHIP

cortical actin

processes 35%

NP

NP

NP

4

FAM40B

cortical actin

processes

elongated 65%

NP

NP

FNBP3

cortical actin, stress fibres

processes

elongated 65%

larger

4%

FMNL3

NP

processes

elongated 80%

larger

NP

Rac1

stress fibres

NP

elongated 65%

larger

NP

LIMD1

NP

processes

elongated 50%

NP

NP

SH3KBP1

NP

NP

elongated 90%

NP

NP

RhoA

strong patches

NP

elongated 70%

NP

NP

SH3D19

spots

NP

very elongated 90%

NP

NP

5

ZMYM4

spots

processes

very elongated 95%

NP

NP

WAVE3

spots

processes

very elongated 90%

NP

NP

LARP4

NP

processes

very elongated 45%

NP

NP

DIAPH1

NP

processes

very elongated 90%

larger

NP

RhoU

NP

NP

very elongated 80%

NP

NP

6

CAPZB

strong patches 90%, cortical actin

NP

NP

NP

NP

CFL1

strong patches 80%

NP

NP

NP

31%

PDZK8

strong patches 45%

NP

NP

NP

2%

7

ZMYM6

rough edge

NP

rounded 65%

NP

NP

Cdc42

rough edge

processes

rounded 70%

NP

NP

WTIP

NP

NP

rounded 95%

NP

NP

BRWD1

NP

NP

rounded 80%

smaller

NP

BRWD3

NP

NP

rounded 80%

smaller

NP

HYPC

low 65%, smooth

NP

NP

NP

NP

EPB41L4A

NP

NP

NP

NP

NP


Phenotypes were classified three days after transfection with siRNA pools targeting each of the 26 PMMs or 8 known regulators of the actin cytoskeleton (italics). Cells were fixed and stained for F-actin, microtubules, and DNA (as in Figure 1). Three to four different images from two different screens were analysed; total number of cells analysed/condition ≥ 68. The most prominent phenotype for each gene is highlighted in bold text and the percent of cells showing this phenotype is shown in brackets (see Figure 1, Supplementary. Figure 2 for examples of each phenotype). Definitions of actin phenotypes: stress fibres, multiple thick straight F-actin cables/cell; cortical actin, increased F-actin on regions of the plasma membrane; spots, small bright regions of F-actin; strong patches, large bright regions of F-actin; rough edge, free edges of cells are wavy or indented; smooth, F-actin levels similar throughout cytoplasm; low, lower F-actin levels than control cells. Microtubule phenotype: processes, long narrow membrane extensions containing microtubules. Definitions of cell shape: larger, spread area increased compared to control cells; elongated, cell body length ≥ 2 × width; very elongated, cell body length ≥ 3 × width; rounded, length ~ = width. NP, no phenotype.

Bai et al. BMC Biology 2011 9:54   doi:10.1186/1741-7007-9-54

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