Anti c myc tag

The cell lysates were subjected to SDS-PAGE, then the signaling properties were analyzed by western blotting. The list of rabbit primary antibodies used in this study is described as follows: anti phospho-STAT1 (Y701)(Cell Signaling Technology, Danvers, MA), anti STAT1 (Cell Signaling Technology), anti phospho-STAT3 (Y705) (Cell Signaling Technology), anti STAT3 (Santa Cruz Biotechnology, Santa Cruz, CA), anti phospho-STAT5 (Y694) (Cell Signaling Technology), anti STAT5 (Santa Cruz Biotechnology), anti HA tag (BETHYL), anti V5 tag (Millipore, Burling, MA), anti c-myc tag (BETHYL, Montgomery, TX), and anti GAPDH (Santa Cruz Biotechnology). As a secondary antibody, horseradish peroxidase-conjugated anti rabbit IgG (Thermo Fisher Scientific) was commonly used.

The protocol for signaling analysis was described previously^{14 (link)}. In brief, the stimulated cells (10⁶ cells) were washed immediately by ice-cold PBS containing 2 mM Na₃VO₄ twice, lysed with 100 μl of lysis buffer (20 mM HEPES (pH 7.5), 150 mM NaCl, 10% glycerol, 1% Triton X-100, 1.5 mM MgCl₂, 1 mM EGTA, 10 μg/ml aprotinin, 10 μg/ml leupeptin), and incubated on ice for 10 min. After centrifugation at 21,500 g for 10 min at 4°C, the supernatant was mixed with 33 μl of 4 x Laemmli’s buffer and boiled. The cell lysates were loaded to SDS-PAGE to analyze the signaling properties by western blotting. Following is the list of the rabbit primary antibodies used in this study: anti phospho-STAT1 (Y701)(Cell Signaling Technology, Danvers, MA), anti STAT1 (Cell Signaling Technology, Danvers, MA), anti phospho-STAT3 (Y705) (Cell Signaling Technology), anti STAT3 (Santa Cruz Biotechnology, Santa Cruz, CA), anti phosph-STAT5 (Y694) (Cell Signaling Technology), anti STAT5 (Santa Cruz Biotechnology), anti GAPDH (Santa Cruz Biotechnology), anti c-myc tag (BETHYL, Montgomery, TX). Horseradish peroxidase-conjugated anti rabbit IgG (Thermo Fisher Scientific) was used as a secondary antibody.