Table 1. Drosophila in vivo screening for phagocyte factors protective against L. amazonensis infection.
Gene knockdown was targeted to hemocytes by crossing a fly line carrying the Hml(Δ)-Gal4-driver to UAS-RNAi-expressing flies. Croquemort and draper mutant lines were also analyzed, as indicated. The 3rd column presents the percentage of assays where the survival of the experimentally-infected flies was significantly reduced compared to PBS-injected RNAi flies and to control amastigote-infected flies (p<0.005), the total number of independent survival assays are presented in parentheses. Fly lines that were susceptible in more than 60% of the assays were considered hits (bold). The hyper-susceptibility of fly lines deficient in Rac2, ftz-f1 and ß’COP, demonstrates that the approach identifies known phagocytic factors. The comparison of the curves was performed using a Log-rank (Mantel-Cox) test. Representative survival curves of susceptible flies are presented in Fig 3.
| Gene name or symbol | Protein family or feature | % assays with reduced survival (n) |
|---|---|---|
| croquemort | Scavenger receptor Class B | 71 (7) |
| croquemort (mutant) | Scavenger receptor Class B | 75 (4) |
| CG31741 | Scavenger receptor Class B | 62 (8) |
| CG10345 | Scavenger receptor Class B | 71 (7) |
| debris buster | Scavenger receptor Class B | 25 (4) |
| CG2736 | Scavenger receptor Class B | 33 (3) |
| epithelial membrane protein | Scavenger receptor Class B | 33 (3) |
| peste | Scavenger receptor Class B | 0 (2) |
| CG7227 | Scavenger receptor Class B | 0 (7) |
| santa-maria | Scavenger receptor Class B | 0 (2) |
| Sensory neuron membrane protein 1 | Scavenger receptor Class B | 0 (3) |
| CG3829 | Scavenger receptor Class B | 0 (2) |
| Scavenger receptor class C, type III | Scavenger receptor Class C | 75 (8) |
| Scavenger receptor class C, type IV | Scavenger receptor Class C | 67 (6) |
| Scavenger receptor class C, type II | Scavenger receptor Class C | 25 (8) |
| Nimrod A | Scavenger receptor Class F-like | 33 (3) |
| Nimrod B1 | Scavenger receptor Class F-like | 0 (3) |
| Nimrod B2 | Scavenger receptor Class F-like | 0 (2) |
| Nimrod B5 | Scavenger receptor Class F-like | 0 (2) |
| Nimrod C1 | Scavenger receptor Class F-like | 33 (3) |
| Nimrod C2 | Scavenger receptor Class F-like | 33 (3) |
| Nimrod C3 | Scavenger receptor Class F-like | 63 (8) |
| Nimrod C4 | Scavenger receptor Class F-like | 33 (3) |
| eater | Scavenger receptor Class F-like | 0 (3) |
| slowdown | Scavenger receptor Class F-like | 0 (2) |
| draper (mutant) | Scavenger receptor Class F-like | 0 (2) |
| ftz transcription factor 1 | Nuclear hormone receptor | 100 (3) |
| Rac2 | Ras GTPase | 100 (3) |
| Coat Protein (coatomer) β' | WD repeats | 75 (4) |
| phagocyte signaling impaired | Tetratricopeptide domains repeat | 33 (3) |
| SCAR | Actin-binding protein | 0 (2) |
| Rab5 | Ras GTPase | 0 (2) |