Proteins for western blotting were extracted from the brain and SG or the brain--SG complexes of pupae, quantified by the Bradford method [ 49], and stored at -80°C.
Genomic DNA for Southern blotting was extracted from three cell lines and the sections of the frozen material of six patient samples using standard protocols.
For western blotting, proteins were extracted from tissues or cultured cells using RIPA buffer containing phenylmethanesulfonylfluoride (Beyotime, Nantong, China).
Concerning Western blot analysis, proteins were extracted from jejunal mucosa scrapings as described above.
For western blotting, total proteins were extracted from cells cultivated in appropriate selective medium, submitted to SDS-PAGE electrophoresis, and HA-tagged Imp3 proteins were revealed with the anti-HA 3F10 antibody from Roche, as described in [39].
For western blotting analysis, proteins were extracted from cultured cardiomyocytes using Trizol reagent.
For Western blot analysis, proteins were extracted from normal and tumour tissues in five representative breast cancer cases.
Proteins for western blot were extracted either from Trizol samples according to the manufacturer's instructions, or from virally infected tadpoles homogenised in 10 mM Tris with 100 µL sterile 1.0 mm zirconia silicone beads (BioSpec Products).
For northern blot assay, small RNAs were extracted from HEK293T or HeLa cells by first preparing total RNA with Tri-reagent (MRC inc).
For western blot analysis, total proteins were extracted from 1-month-old leaves in Laemmli buffer.
