Plate Format:

384

Origin:	Human
Cell Medium:	DMEM (10%FBS, P/S)
Cell Seeding Density Per Well:	600cells
Cell Culture Conditions:	10%CO2, 37degree

Protocol:	Mix 2.5ul 375nM siRNA in HBSS and 0.0075ul/well Interferin-HTS in 2.5ul OptiMEM. Incubate for 15mins, then add cells in 45ul.
Delivery Reagent:	Interferin-HTS
Delivery Reagent Supplier:	Polyplus
Delivery Reagent Concentration:	0.0075ul/well
Silencing Reagent Concentration:	18.75nM
Silencing Reagent Supplier:	Dharmacon
Silencing Reagent Origin:	HTS lab

BAF A1 treatment to induce Autophagy
Treatment:After 72hrs of transfection, treat the cells with BAF A1 (50nM) for 6hours to induce antophagy by adding 25ul of 150nM BAF A1 media into 45ul culture.
Compounds:Bafilomycin A1
Compound Concentration:50nM
Treatment Time:6 hours

Description:Fix with 4%PFA by top-up 25ul 16%PFA
Fixation Compounds:16%PFA
Compound Concentration:4%PFA
Treatment Time:30min

Secondary Antibody - Alexa Fluor 488 donkey anti-mouse IgG secondary antibody
Supplier:Molecular Probes
Cat No:A-21202
Batch No:436938
Dilution:1/1000
Incubation Time:1 hour
Secondary Antibody - 647 donkey anti-rabbit IgG
Supplier:Invitrogen
Cat No:A31573
Batch No:1874788
Dilution:1:1000
Incubation Time:1 hour
Primary Antibody - LC3 mouse
Supplier:Nanotool
Dilution:1:100
Incubation Time:2hr
Primary Antibody - P62 rabbit
Supplier:Abcam
Dilution:1:6000
Incubation Time:2hr

Nuclear Staining:DAPI
Nuclear Staining Concentration:1ug/ml
Nuclear Staining Incubation Time:1 hour