Fig. 3: Galectin-3 senses LPS to activate mTORC1.

a HEK-293T cells were infected with lentiviruses carrying shGFP or shGal3, starved of amino acids for 50 min, and replenished with amino acids for 10 min for western blotting analysis of the level or phosphorylation of the indicated proteins. Each experiment was repeated three times with similar results. b HEK-293T cells were starved of amino acids and glucose for 1 h followed by stimulation with LPS (1 μg/ml) for 5 h in the presence or absence of the mTOR inhibitor Torin1 (250 nM) or galectin-3 inhibitor GB1107 (5 μM) for western blotting analysis of the level or phosphorylation of the indicated proteins. Each experiment was repeated three times with similar results. c HEK-293T cells were infected with lentiviruses carrying shGFP or shGal3, or HA-Vector or HA-Gal3 treated with or without GB1107 (5 μM) or Torin1 (250 nM), and starved of amino acids and glucose for 1 h followed by stimulation with LPS (1 μg/ml) for 5 h for western blotting analysis of the level or phosphorylation of the indicated proteins. Each experiment was repeated three times with similar results. d HEK-293T cells were transfected with the indicated plasmids, starved of amino acids and glucose for 1 h, and stimulated with LPS (1 μg/ml) for 5 h. Cellular lysates were immunoprecipitated with anti-FLAG followed by immunoblotting with antibodies against the indicated proteins. Each experiment was repeated three times with similar results.