Comprehensive Western Blotting Analysis

Western blotting was performed as described as previously depicted [15 (link)]. And immunoblotted with the following antibodies: anti-mouse SLC3A2 (1:1000, Santa, sc-376,815, USA), anti-mouse mTOR (1:1000, Santa, sc-517,464, USA), anti-mouse p-mTOR (1:1000, Santa, sc-293,133, USA), anti-mouse P70S6K (1:1000, Santa, sc-8418, USA), anti-mouse p-P70S6K (1:1000, Santa, sc-8416, USA), anti-mouse FTH1 (1:1000, Santa, sc-376,594, USA), anti-mouse TFR1 (1:1000, Santa, sc-32,272, USA), anti-Rabbit HO-1 (1:1000, abcam, ab52947, England), anti-mouse GPX4 (1:1000, Santa, sc-166,120, USA), and anti-mouse GAPDH (1:1000, Santa, sc-47,724, USA). Then, the PVDF membranes were washed and secondary antibodies were applied 1:5000 for 1 h at room temperature, the immunoreactions were visualized with chemiluminescent ECL reagent.