MK-571 was purchased from Merck Sharp (Kirkland, Canada) Proteins for Western blot analysis were extracted by lysing cells with sample buffer containing 0.125 M Tris-HCl, 2% SDS, 10% glycerol and 0.001% bromophenol blue.
For Western blotting, EMV proteins were extracted by boiling 5-10 min in Laemmli buffer (BioRad, Hercules, CA, USA) containing fresh 5%% beta-mercaptoethanol (Sigma-Aldrich, St . Louis MO, USA).
For western blot analysis, proteins were extracted by scraping the NPCs in homogenate buffer (320 mM sucrose, 1 mM EDTA, 50 mM Tris-HCl, 1 mM PMSF and 1 × proteinase inhibitor).
For northern blots, RNA was extracted by resuspending cells in 50 mM Tris HCl pH 7.5, 10 mM EDTA pH 8, 100 mM NaCl, 1% SDS, lysing by the addition of phenol chloroform 5 1, acid washed beads and vortexing for 30 min at 4°C.
Western blotting and immunohistochemistry Proteins were extracted, quantified and separated by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membranes using previously described methods [ 31].
Nuclear and cytoplasmic proteins were extracted and assayed by western blotting with the indicated antibodies.
Total kidney proteins were extracted and analyzed by Western blotting.
Coprecipitated RNAs were extracted and analyzed by northern blotting.
The proteins of the transfected cells were extracted for Western blot by using the anti-GFP antibody and KIAA0101 rabbit antibody to test the expression of EGFP and KIAA0101.
Lysates were extracted and subsequently analyzed by western blotting.
