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. 2021 Aug 27;22(17):9289. doi: 10.3390/ijms22179289

Figure 3.

Figure 3

(20S)G-Rh2 inhibited STAT3 activation by targeting Anxa2. (A) Thermal shift assay was performed with purified Anxa2 and (20S)G-Rh2 gradient at 55 °C, and determined by immuno-blotting. (B) Cellular thermal shift assay was performed in HepG2 cells with indicated temperature gradient and determined by immuno-blotting. (C–E) HepG2 cells were treated with Src inhibitor (Saracatinib), Src activator (Src inhibitor 3) or (20S)G-Rh2, or under co-treatment. Immuno-precipitation with anti-Anxa2 antibody (C) and protein level (D) were determined by immuno-blotting. STAT3 activation was determined via reporter gene assay (E). (F–H) HepG2 cells were transfected with the constitutively activated mutant of Src and treated with (20S)G-Rh2. Immuno-precipitation with anti-Anxa2 antibody (F) and protein level (G) were determined by immuno-blotting. STAT3 activation was determined via reporter gene assay (H). All experiments were performed with triple replicant. Significance analysis was shown as ns (p > 0.05) and *** (p < 0.001).