l
yw hsp-flipase; Sp/CyO; FRT82B Gal80/TM6b.
Lines for making ApcRas clones (benign CRC)
l
yw hsp-flipase; esg Gal4, UAS-GFP, UAS-RasV12/CyO; UASLuciferase FRT82B Gal80/TM6b.
l
yw hsp-flipase; Sp/CyO; FRT82B Apc2N175KApcQ8/TM6b.
Lines for making ApcRasSna clones (metastatic CRC)
l
yw hsp-flipase; esg Gal4, UAS-GFP, UAS-RasV12/CyO; UASLucifersase FRT82B Gal80/TM6b.
l
yw hsp-flipase; UAS-Sna/CyO; FRT82B Apc2N175KApcQ8/
TM6b.
Apc2
N175K is a loss-of-function allele, Apc
Q8 is a null allele,
UAS-Ras
V12 is a gain-of-function transgene, and UAS-Sna is a
wild-type transgene. Stocks were obtained from Bloomington
Stock Center, except UAS-Sna (kind gift of Kumar lab).
2.2 Equipment
and Reagents
1. Parafilm.
2. A fluorescence stereo microscope.
3. Two pairs of sharp forceps.
4. Microscope slides.
5. Phosphate-Buffered Saline (PBS).
6. Oil 10 S, VOLTALEF.
7. Electron microscope grade paraformaldehyde.
8. Bovine serum albumin (BSA).
9. Mounting medium such as Vectashield or Fluoromount.
10. Triton X-100.
11. Anti-GFP antibody.
12. Phalloidin-TRITC.
13. Confocal microscope.
14. Tungsten needle.
15. Luciferase reporter assay kit.
16. A microplate reader that can detect luminescence.
17. 96-Well microplates.
2.3 Image Analysis
For image analysis, we recommend using ImageJ Fiji distribution
(https://imagej.net/Fiji/Downloads). In principle Fiji should run
on all major computer platforms, including Microsoft Windows,
Mac OS, and Linux.
Studying Metastatic Cancer Progression in vivo
163
yw hsp-flipase; Sp/CyO; FRT82B Gal80/TM6b.
Lines for making ApcRas clones (benign CRC)
l
yw hsp-flipase; esg Gal4, UAS-GFP, UAS-RasV12/CyO; UASLuciferase FRT82B Gal80/TM6b.
l
yw hsp-flipase; Sp/CyO; FRT82B Apc2N175KApcQ8/TM6b.
Lines for making ApcRasSna clones (metastatic CRC)
l
yw hsp-flipase; esg Gal4, UAS-GFP, UAS-RasV12/CyO; UASLucifersase FRT82B Gal80/TM6b.
l
yw hsp-flipase; UAS-Sna/CyO; FRT82B Apc2N175KApcQ8/
TM6b.
Apc2
N175K is a loss-of-function allele, Apc
Q8 is a null allele,
UAS-Ras
V12 is a gain-of-function transgene, and UAS-Sna is a
wild-type transgene. Stocks were obtained from Bloomington
Stock Center, except UAS-Sna (kind gift of Kumar lab).
2.2 Equipment
and Reagents
1. Parafilm.
2. A fluorescence stereo microscope.
3. Two pairs of sharp forceps.
4. Microscope slides.
5. Phosphate-Buffered Saline (PBS).
6. Oil 10 S, VOLTALEF.
7. Electron microscope grade paraformaldehyde.
8. Bovine serum albumin (BSA).
9. Mounting medium such as Vectashield or Fluoromount.
10. Triton X-100.
11. Anti-GFP antibody.
12. Phalloidin-TRITC.
13. Confocal microscope.
14. Tungsten needle.
15. Luciferase reporter assay kit.
16. A microplate reader that can detect luminescence.
17. 96-Well microplates.
2.3 Image Analysis
For image analysis, we recommend using ImageJ Fiji distribution
(https://imagej.net/Fiji/Downloads). In principle Fiji should run
on all major computer platforms, including Microsoft Windows,
Mac OS, and Linux.
Studying Metastatic Cancer Progression in vivo
163