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.