Real-time PCR and western blot assay

Total RNA was isolated by using TRIzol reagent and additional phenol. Mature miR-150 expression was detected using a quantitative real-time PCR assay with miR-150-specific primers and a TaqMan miR-150 probe. The relative amount of SRCIN1 mRNA was normalized to that of GAPDH.

For western blot assay, cell lysates were separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel and transferred to a nitrocellulose membrane. The membranes were probed with corresponding antibodies. The protein levels were normalized with β-actin.