Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference |
Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Homo sapiens) | Raf1 | PMID:24667437 | NCBI Gene ID: 207 | |
| Gene (Homo sapiens) | AKT | PMID:27082641 | NCBI Gene ID: 5894 | |
| Strain, strain background (D. melanogaster) | 19–12-Gal4 | PMID:21068723 | FLYB: FBti0148308 | FlyBase symbol: P{GAL4}19–12 |
| Strain, strain background (D. melanogaster) | repo-Gal80 | PMID:19091965 | FLYB: FBtp0067904 | Flybase symbol: P{repo-GAL80.L} |
| Strain, strain background (D. melanogaster) | ppk-CD4-tdGFP | PMID:21606367 | BDSC: 35842 FLYB: FBti0143429 RRID:BDSC_35842 |
Flybase symbol: P{ppk-CD4-tdGFP}1b |
| Strain, strain background (D. melanogaster) | ppk-Gal4 | PMID:21606367 | BDSC: 32079 FLYB: FBti0131208 RRID:BDSC_32079 |
Flybase symbol: P{ppk-GAL4.G}3 |
| Strain, strain background (D. melanogaster) | UAS-optoRaf | This paper | Transgenic fly with inducible expression of optoRaf, Dr. Yuanquan Song | |
| Strain, strain background (D. melanogaster) | UAS-optoAKT | This paper | Transgenic fly with inducible expression of optoAKT, Dr. Yuanquan Song | |
| Cell line (Homo sapiens) | HEK293T | PMID:32277988 | Dr. Lin-Feng Chen | |
| Cell line (Rattus norvegicus) | PC12 | PMID:22206868 | Dr. Tobias Meyer | |
| Cell line (Mesocricetus auratus) | BHK-21 | PMID:26080442 | Dr. Steven Chu | |
| Transfected construct (Homo sapiens) | pEGFP N1 CRY2-mCh-Raf1-P2A-CIBNx2-EGFP-CAAX | This paper | OptoRaf, CIBN-CRY2 based Raf membrane translocation system. Dr. Kai Zhang | |
| Transfected construct (Homo sapiens) | pEGFP N1 CRY2-mCh-Raf1 S338A-P2A-CIBNx2-EGFP-CAAX | This paper | OptoRaf S338A CIBN-CRY2 based Raf S338A membrane translocation system. Dr. Kai Zhang | |
| Transfected construct (Homo sapiens) | pEGFP N1 CRY2-mCh-AKT-P2A-CIBNx2-EGFP-CAAX | This paper | OptoAKT, CIBN-CRY2 based AKT membrane translocation system. Dr. Kai Zhang | |
| Transfected construct (Arabidopsis thaliana) | pEGFP C1 CIBN-EGFP-CAAX | PMID:21037589 | Dr. Chandra Tucker | |
| Transfected construct (Homo sapiens) | pEGFP C1 Raf1-EGFP-CaaX | PMID:24667437 | A constitutively active form of Raf1. Dr. Kai Zhang | |
| Transfected construct (Rattus norvegicus) | pEGFP N1 ERK2-EGFP | PMID:24667437 | Fluorescently label ERK2 for nuclear translocation assay. Dr. Kai Zhang | |
| Transfected construct (Homo sapiens) | pEGFP N1 FOXO3-EGFP | PMID:10102273 | Fluorescently labeled FOXO3 for nuclear export assay. Dr. Kai Zhang | |
| Antibody | Anti-Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204), rabbit monoclonal | Cell Signaling Technology | Cat# 4370T RRID:AB_2315112 |
IF (1:400) |
| Antibody | Anti-Phospho-Drosophila p70 S6 Kinase (Thr398), rabbit polyclonal | Cell Signaling Technology | Cat# 9209S RRID:AB_2269804 |
IF (1:100) |
| Antibody | Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204), rabbit polyclonal | Cell Signaling Technology | Cat# 9101S RRID:AB_331646 |
WB (1:1000) |
| Antibody | p44/42 MAPK (Erk1/2), rabbit polyclonal | Cell Signaling Technology | Cat# 9102S RRID:AB_330744 |
WB (1:1000) |
| Antibody | c-Raf, rabbit polyclonal | Cell Signaling Technology | Cat# 9422S RRID:AB_390808 |
WB (1:1000) |
| Antibody | Phospho-c-Raf (Ser338) (56A6), rabbit monoclonal | Cell Signaling Technology | Cat# 9427S RRID:AB_2067317 |
WB (1:1000) |
| Antibody | Phospho-Akt (Ser473) (D9E) XP, rabbit monoclonal | Cell Signaling Technology | Cat# 4060S RRID:AB_2315049 |
WB (1:1000) |
| Antibody | Phospho-Akt (Thr308) (244F9), rabbit monoclonal | Cell Signaling Technology | Cat# 4056S RRID:AB_331163 |
WB (1:1000) |
| Antibody | Akt (pan) (C67E7), rabbit monoclonal | Cell Signaling Technology | Cat# 4691S RRID:AB_915783 |
WB (1:1000) |
| Antibody | PLCγ1, rabbit polyclonal | Cell Signaling Technology | Cat# 2822S RRID:AB_2163702 |
WB (1:1000) |
| Antibody | Phospho-PLCγ1 (Tyr783), rabbit polyclonal | Cell Signaling Technology | Cat# 2821S RRID:AB_330855 |
WB (1:1000) |
| Antibody | GAPDH (14C10) Rabbit monoclonal | Cell Signaling Technology | Cat# 2118S RRID:AB_561053 |
WB (1:1000) |
| Antibody | Anti-rabbit IgG, HRP-linked Antibody (Goat anti-rabbit) |
Cell Signaling Technology | Cat# 7074S RRID:AB_2099233 |
WB (1:1000) |
| Antibody | fluorescence-conjugated secondary antibodies (Donkey anti-rabbit 647) | Jackson ImmunoResearch | Cat# 711-605-152 RRID:AB_2492288 |
WB (1:1000) |
| Recombinant DNA reagent | pACU2 (plasmid) | PMID:21606367 | RRID:Addgene_31223 | |
| Chemical compound, drug | VECTASHIELD Antifade Mounting Medium (with DAPI) | Vector Laboratories | Cat# H-1200 | |
| Software, algorithm | ImageJ (Fiji) | http://fiji.sc | RRID:SCR_002285 | |
| Software, algorithm | NeuronStudio | https://biii.eu/neuronstudio | RRID:SCR_013798 |