Western Blotting Analysis of Cell Signaling

Western blotting analysis used in this study was performed as previously described (24 (link)). Antibodies against human HK2, cyclin A1, p27, p-Raf-1, MEK1/2, p-MEK1/2, ERK1/2, c-myc, GAPDH were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-p-ERK1/2 was purchased from CST (Littleton, CO, USA). The horseradish peroxidase-conjugated anti-rabbit or anti-mouse IgG was purchased from Thermo Fisher Scientific (New York, NY, USA). The antibodies used were as follows: anti-HK2 (1:500 dilution, sc-374091, Santa Cruz, USA), anti-cyclin A1 (1:1,000 dilution, sc-239, Santa Cruz, USA), anti-p27 (1:300 dilution, sc-1641, Santa Cruz, USA), anti-MEK1/2 (1:500 dilution, sc-81504, Santa Cruz, USA), anti-p-MEK1/2 (1:500 dilution, sc-81503, Santa Cruz, USA), anti-ERK1/2 (1:500 dilution, sc-135900, Santa Cruz, USA), anti- p-Raf-1 (1:500 dilution, sc-271919, Santa Cruz, USA), anti-c-myc (1:500 dilution, sc-40, Santa Cruz, USA), anti-GAPDH (1:500 dilution, sc-47724, Santa Cruz, USA), anti-p-ERK1/2 (1:200 dilution, #4370, Cell Signaling Technology). GAPDH was used as the control and for quantification.