EGCG induces SASP in human HSCs cell line in vitro. TWNT4 cells were treated with different concentration of EGCG (0, 2, 5, 10, 20, 40, 50, and 100 μmol/L) for 72 hours. A, Representative bright field pictures of TWNT4 cells treated with 20, 50, and 100 μmol/L of EGCG. B, MTT cell proliferation assay in TWNT4 cells, 72 hours after treatment with EGCG. C and D, Immunofluorescence staining of Ki67 (red) and DAPI (blue) on TWNT4 cells after 72 hours treatment with 20 μmol/L EGCG. Untreated cells were considered as control, 2,000 cells were counted in each group; ***, P < 0.001 compared with control. E and F, SA-β-gal staining (turquoise blue) of TWNT4 cells after 72 hours treatment with 20 μmol/L EGCG. Untreated cells were considered as control, 2,000 cells were counted in each group; **, P < 0.01 compared with control. G-J, RNA expression of senescence and SASP markers in TWNT4 cells after 72 hours treatment with 20 μmol/L EGCG. Untreated cells were considered as control. N = 4 replicates (*, P < 0.05; **, P < 0.01; ***, P < 0.001).