Western blotting analysis was carried out in accordance with the procedures outlined in the previous section (Mo et al., 2020 (link)). SDS-PAGE and polyvinylidene fluoride membranes were used to separate the protein lysates. The membranes were blocked for 1 h in Tris-buffered saline containing 0.1% Tween 20 before being probed with specific antibodies. Primary antibodies included p-AKTThr308, t-AKT, p-mTOR, p-RPS6, SREBP1, FASN, ACC, HK2, and PKM2. Anti-rabbit/mouse IgG/HRP (secondary antibody) was added following incubation with each primary antibody. HRP-conjugated β-actin antibody from Proteintech (Wuhan, China) was used as an internal standard. Using SuperSignal West Pico Chemiluminescent Substrate (Pierce Chemical, Ridgewood, New York, United States) and a chemiluminescence imager, immunoblotting was performed and photographed (G: BOX Chemi XRQ; Syngene, Cambridge, United Kingdom).
The IHC assay was performed as previously described (Hu et al., 2016 (link)). Specific primary antibodies used in immunohistochemical staining are listed inSupplementary Table S1 .
The IHC assay was performed as previously described (Hu et al., 2016 (link)). Specific primary antibodies used in immunohistochemical staining are listed in
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