The Western blotting analysis was performed according to the protocol we have reported [28 (link)–30 (link)]. The anti-HHLA2 (1:2000; Abcam, MA, USA), the anti-GAPDH (1:4000, Sigma, St. Louis, MO, USA), and the HRP-labeled goat anti-mouse/rabbit secondary antibody (1:6000, Sigma Aldrich, St. Louis, MO, USA) were used, and the immunoreaction was visualized using an enhanced chemiluminescence detection kit (Thermo Fisher, MA, USA) and exposed to X-ray film, and band densities were quantified by densitometry with a video documentation system (Gel Doc 2000, Bio-Rad).
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