Anti mouse hrp linked

Pre-stained markers and protein samples with equal amounts of total protein were separated by electrophoresis on 8 % SDS-PAGE gels and transferred to polyvinylidene difluoride membranes (Millipore, USA). The membranes were blocked with TBST containing 5 % non-fat milk. The procedure was followed by overnight incubation with a monoclonal antibody to MMP2 (1:1000) or MMP9 (1:500; Abcam, UK) at 4 °C, six washes with 0.01 % Tween-TBS buffer, and a 2 h incubation with a secondary antibody (anti-mouse HRP-linked, 1:7000; Abcam, UK). GAPDH (1:100; Boster, China) was used as control for equal protein loading. The band intensities were quantitated using the Image Pro Plus 5.1 software.

Antibodies used were from Cell Signaling (Danvers, MA, USA) against DYRK1A, P107, P16, p-RB, RB, p-CycD1, CycD1, CycB1, p-CDK1, Ki67, GAPDH, anti-rabbit HRP-linked, anti-mouse HRP-linked; from Abcam (Cambridge, UK) against CDK1, p-CDK1, CDC23, LIN52; and from Santa Cruz Biotechnology (Dallas, TX, USA) against LIN37, DYRK1A, LIN9, P130.